Crystalline forms and processes for the preparation of cannabinoid receptor modulators

ABSTRACT

The present invention relates to crystalline forms of (1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylic acid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1) and pharmaceutical compositions thereof that modulate the activity of the cannabinoid CB 2  receptor and are therefore useful in the treatment of CB 2  receptor-mediated disorders, for example, osteoarthritis; pain; hyperalgesia; allodynia; inflammatory hyperalgesia; neuropathic hyperalgesia; acute nociception; osteoporosis; multiple sclerosis-associated spasticity; autoimmune disorders; allergic reactions; CNS inflammation for example; atherosclerosis; undesired immune cell activity, and inflammation associated with a disorder selected from: osteoarthritis, anaphylaxis, Behcet&#39;s disease, graft rejection, vasculitis, gout, spondylitis, viral disease, bacterial disease, lupus, inflammatory bowel disease, autoimmune hepatitis, and type 1 diabetes mellitus; age-related macular degeneration; cough; leukemia; lymphoma; CNS tumors; prostate cancer; Alzheimer&#39;s disease; stroke-induced damage; dementia; amyotrophic lateral sclerosis; and Parkinson&#39;s disease.

FIELD OF THE INVENTION

The present invention relates to crystalline forms of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1) andpharmaceutical compositions thereof that modulate the activity of thecannabinoid CB₂ receptor and are therefore useful in the treatment ofCB₂ receptor-mediated disorders, for example, osteoarthritis; pain, forexample bone and joint pain, muscle pain, dental pain, migraine andother headache pain, inflammatory pain, neuropathic pain, pain thatoccurs as an adverse effect of therapeutics, and pain associated with adisorder selected from: osteoarthritis, cancer, multiple sclerosis,allergic reactions, nephritic syndrome, scleroderma, thyroiditis,diabetic neuropathy, fibromyalgia, HIV related-neuropathy, sciatica, andautoimmune conditions; hyperalgesia; allodynia; inflammatoryhyperalgesia; neuropathic hyperalgesia; acute nociception; osteoporosis;multiple sclerosis-associated spasticity; autoimmune disorders, forexample an autoimmune disorder selected from the group consisting of:multiple sclerosis, Guillan-Barré syndrome, polyradiculoneuropathy,chronic inflammatory demyelination, rheumatoid arthritis, psoriaticarthritis, ankylosing spondylarthritis, and reactive arthritis; allergicreactions, for example, an allergic reaction associated with a disorderselected from: atopic dermatitis, pruritis, urticaria, asthma,conjunctivitis, allergic rhinitis, and anaphylaxis; CNS inflammation forexample, CNS inflammation associated with a disorder selected from:Alzheimer's disease, stroke, dementia, amyotrophic lateral sclerosis,and human immunodeficiency virus; atherosclerosis; undesired immune cellactivity, and inflammation associated with a disorder selected from:osteoarthritis, anaphylaxis, Behcet's disease, graft rejection,vasculitis, gout, spondylitis, viral disease, bacterial disease, lupus,inflammatory bowel disease, autoimmune hepatitis, and type 1 diabetesmellitus; age-related macular degeneration; cough; leukemia; lymphoma;CNS tumors; prostate cancer; Alzheimer's disease; stroke-induced damage;dementia; amyotrophic lateral sclerosis, and Parkinson's disease. Thepresent invention further relates to processes and intermediates usefulin the preparation crystalline forms and solvates of Compound 1 andpharmaceutical compositions thereof.

BACKGROUND OF THE INVENTION

Cannabinoids are a group of extracellular signaling molecules that arefound in both plants and animals. Signals from these molecules aremediated in animals by two G-protein coupled receptors, CannabinoidReceptor 1 (CB₁) and Cannabinoid Receptor 2 (CB₂). CB₁ is expressed mostabundantly in the neurons of the CNS but is also present at lowerconcentrations in a variety of peripheral tissues and cells (Matsuda, L.A. et al. (1990) Nature 346:561-564). In contrast, CB₂ is expressedpredominantly, although not exclusively, in non-neural tissues, e.g. inhematopoietic cells, endothelial cells, osteoblasts, osteoclasts, theendocrine pancreas, and cancerous cell lines (Munro, S. et al. (1993)Nature 365:61-65; and as reviewed in Pacher, P. et al. (2006) Pharmacol.Rev. 58(3): 389-462). As such, CB₁ is believed to be primarilyresponsible for mediating the psychotropic effects of cannabinoids onthe body, whereas CB₂ is believed to be primarily responsible for mostof their non-neural effects.

SUMMARY OF THE INVENTION

One aspect of the present invention relates to crystalline forms of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1):

One aspect of the present invention relates to an anhydrous crystallineform of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1).

One aspect of the present invention relates to processes for preparingan anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide comprising thesteps of:

-   -   1) crystallizing        (1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylic        acid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide from a        crystallizing mixture to obtain a crystalline form of        (1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylic        acid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide in the        crystallizing mixture, wherein the crystallizing mixture        comprises acetonitrile and water; and    -   2) isolating the crystalline form of        (1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylic        acid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide from the        crystallizing mixture to obtain the anhydrous crystalline form        of        (1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylic        acid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide.

One aspect of the present invention relates to an anhydrous crystallineform of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide prepared by aprocess as described herein.

One aspect of the present invention relates to compositions comprisingan anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide as describedherein.

One aspect of the present invention relates to compositions comprisingan anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide as describedherein, and a pharmaceutically acceptable carrier.

One aspect of the present invention relates to processes of making acomposition comprising mixing an anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide as describedherein, with a pharmaceutically acceptable carrier.

One aspect of the present invention relates to methods for the treatmentof a cannabinoid receptor-mediated disorder in an individual, comprisingadministering to the individual in need thereof, a therapeuticallyeffective amount of an anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide as described hereinor a pharmaceutical composition thereof.

One aspect of the present invention relates to methods for the treatmentof a CB₂ receptor-mediated disorder in an individual, comprisingadministering to the individual in need thereof, a therapeuticallyeffective amount of an anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide as described hereinor a pharmaceutical composition thereof.

One aspect of the present invention relates to the use of an anhydrouscrystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide as describedherein, in the manufacture of a medicament for the treatment of acannabinoid receptor-mediated disorder.

One aspect of the present invention relates to the use of an anhydrouscrystalline form of (laS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide as describedherein, in the manufacture of a medicament for the treatment of a CB₂receptor-mediated disorder.

One aspect of the present invention relates to an anhydrous crystallineform of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide as describedherein, for use in a method of treatment of the human or animal body bytherapy.

One aspect of the present invention relates to an anhydrous crystallineform of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-anide as describedherein, for use in a method of treatment of a cannabinoidreceptor-mediated disorder.

One aspect of the present invention relates to an anhydrous crystallineform of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide as describedherein, for use in a method of treatment of a CB₂ receptor-mediateddisorder.

One aspect of the present invention relates to acetone solvates of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide.

One aspect of the present invention relates to non-selective solvates of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide.

One aspect of the present invention relates to ethyl acetate solvates of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide.

Certain modulators of the cannabinoid receptor are described in PCTapplication PCT/US2010/002360, filed 27 Aug. 2010 (InternationalPublication Number WO2011/025541), and in U.S. provisional applications61/275,506, 61/396,588, and 61/400,146, each of which is incorporatedherein by reference in its entirety.

These and other aspects of the invention disclosed herein will be setforth in greater detail as the patent disclosure proceeds.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a differential scanning calorimetry (DSC) thermogram for asample containing a crystalline form of Compound 1 (CH₂Cl₂ solvate) anda thermogravimetric analysis (TGA) thermogram of a sample containing acrystalline form of Compound 1 (CH₂Cl₂ solvate).

FIG. 2 shows an overlay of a powder X-ray diffraction (PXRD) pattern fora sample containing a crystalline form of Compound 1 (CH₂Cl₂ solvate)obtained from recrystallization using CH₂Cl₂/hexane (Top Trace) and apowder X-ray diffraction (PXRD) pattern for a sample containing acrystalline form of Compound 1 (CH₂Cl₂ solvate) obtained by slurryingnon-solvated Compound 1 in CH₂Cl₂ (Bottom Trace). The PXRD showed thecrystalline solvate obtained from the CH₂Cl₂ slurry is substantiallyindistinguishable from the crystalline solvate resulting fromCH₂Cl₂/hexane recrystallization.

FIG. 3 shows the asymmetric unit for the hemi-CH₂Cl₂ solvate of Compound1 based on single-crystal X-ray diffraction analysis.

FIG. 4 shows a powder X-ray diffraction (PXRD) pattern for a samplecontaining an anhydrous crystalline form of Compound 1.

FIG. 5 shows a differential scanning calorimetry (DSC) thermogram for asample containing anhydrous crystalline form of Compound 1 and athermogravimetric analysis (TGA) thermogram of a sample containinganhydrous crystalline form of Compound 1.

FIG. 6 shows a powder X-ray diffraction (PXRD) pattern for a samplecontaining an anhydrous crystalline form of Compound 1.

FIG. 7 shows a differential scanning calorimetry (DSC) thermogram for asample containing anhydrous crystalline form of Compound 1 and athermogravimetric analysis (TGA) thermogram of a sample containinganhydrous crystalline form of Compound 1.

FIG. 7A shows an adsorption and desorption isotherm, Dyanmic MoistureSorption (DMS), for a sample containing anhydrous crystalline form ofCompound 1.

FIG. 8 shows a powder X-ray diffraction (PXRD) pattern for a samplecontaining a crystalline form of Compound 1 (acetone solvate).

FIG. 9 shows a differential scanning calorimetry (DSC) thermogram for asample containing a crystalline form of Compound 1 (acetone solvate) anda thermogravimetric analysis (TGA) thermogram of a sample containing acrystalline form of Compound 1 (acetone solvate).

FIG. 10 shows a powder X-ray diffraction (PXRD) pattern for a samplecontaining a crystalline form of Compound 1 (non-selective solvate), seeExample 6.

FIG. 11 shows a powder X-ray diffraction (PXRD) pattern for a samplecontaining a crystalline form of Compound 1 (non-selective solvate), seeExample 6.

FIG. 12 shows a powder X-ray diffraction (PXRD) pattern for a samplecontaining a crystalline form of Compound 1 (non-selective solvate), seeExample 6.

FIG. 13 shows a differential scanning calorimetry (DSC) thermogram for asample containing a crystalline form of Compound 1 (non-selectivesolvate) and a thermogravimetric analysis (TGA) thermogram of a samplecontaining a crystalline form of Compound 1 (non-selective solvate), seeExample 6.

FIG. 14 shows a differential scanning calorimetry (DSC) thermogram for asample containing a crystalline form of Compound 1 (non-selectivesolvate) and a thermogravimetric analysis (TGA) thermogram of a samplecontaining a crystalline form of Compound 1 (non-selective solvate), seeExample 6.

FIG. 15 shows a differential scanning calorimetry (DSC) thermogram for asample containing a crystalline form of Compound 1 (non-selectivesolvate) and a thermogravimetric analysis (TGA) thermogram of a samplecontaining a crystalline form of Compound 1 (non-selective solvate), seeExample 6.

FIG. 16 shows a powder X-ray diffraction (PXRD) pattern for a samplecontaining a crystalline form of Compound 1 (ethyl acetate solvate).

FIG. 17 shows a differential scanning calorimetry (DSC) thermogram for asample containing a crystalline form of Compound 1 (ethyl acetatesolvate) and a thermogravimetric analysis (TGA) therlmogram of a samplecontaining a crystalline form of Compound 1 (ethyl acetate solvate).

FIG. 18 shows the effect of Compound 1 in the monosodium iodoacetate(MIA) model of osteoarthritis in rats, see Example 9.

DETAILED DESCRIPTION OF THE INVENTION Definitions

For clarity and consistency, the following definitions will be usedthroughout this patent document.

The term “agonist” refers to a moiety that interacts with and activatesa G-protein-coupled receptor, for instance a cannabinoid receptor, andcan thereby initiate a physiological or pharmacological responsecharacteristic of that receptor. For example, an agonist may activate anintracellular response upon binding to a receptor, or enhance GTPbinding to a membrane.

The term “in need of treatment” and the term “in need thereof” whenreferring to treatment are used interchangeably to mean a judgment madeby a caregiver (e.g. physician, nurse, nurse practitioner, etc. in thecase of humans; veterinarian in the case of animals, including non-humanmammals) that an individual or animal requires or will benefit fromtreatment. This judgment is made based on a variety of factors that arein the realm of a caregiver's expertise, but that includes the knowledgethat the individual or animal is ill, or will become ill, as the resultof a disease, condition or disorder that is treatable by the compoundsof the invention. Accordingly, the compounds of the invention can beused in a protective or preventive manner; or compounds of the inventioncan be used to alleviate, inhibit or ameliorate the disease, conditionor disorder.

The term “individual” refers to any animal, including mammals,preferably mice, rats, other rodents, rabbits, dogs, cats, swine,cattle, sheep, horses, or primates, and most preferably humans.

The term “modulate or modulating” refers to an increase or decrease inthe amount, quality, response or effect of a particular activity,function or molecule.

The term “composition” refers to a compound or crystalline form thereof,including but not limited to, salts, solvates, and hydrates of acompound of the present invention, in combination with at least oneadditional component, such as, a composition obtained/prepared duringsynthesis, preformulation, in-process testing (i.e., TLC, HPLC, NMRsamples), and the like

The term “pharmaceutical composition” refers to a specific compositioncomprising at least one active ingredient; including but not limited to,salts, solvates, and hydrates of compounds of the present invention,whereby the composition is amenable to investigation for a specified,efficacious outcome in a mammal (for example, without limitation, ahuman). Those of ordinary skill in the art will understand andappreciate the techniques appropriate for determining whether an activeingredient has a desired efficacious outcome based upon the needs of theartisan.

The term “therapeutically effective amount” refers to the amount ofactive compound or pharmaceutical agent that elicits the biological ormedicinal response in a tissue, system, animal, individual or human thatis being sought by a researcher, veterinarian, medical doctor or otherclinician or caregiver or by an individual, which includes one or moreof the following:

(1) Preventing the disease, for example, preventing a disease, conditionor disorder in an individual that may be predisposed to the disease,condition or disorder but does not yet experience or display thepathology or symptomatology of the disease;

(2) Inhibiting the disease, for example, inhibiting a disease, conditionor disorder in an individual that is experiencing or displaying thepathology or symptomatology of the disease, condition or disorder (i.e.,arresting further development of the pathology and/or symptomatology);and

(3) Ameliorating the disease, for example, ameliorating a disease,condition or disorder in an individual that is experiencing ordisplaying the pathology or symptomatology of the disease, condition ordisorder (i.e., reversing the pathology and/or symptomatology).

It is appreciated that certain features of the invention, which are, forclarity, described in the context of separate embodiments, may also beprovided in combination in a single embodiment. Conversely, variousfeatures of the invention, which are, for brevity, described in thecontext of a single embodiment, may also be provided separately or inany suitable subcombination. In addition, subcombinations of uses andmedical indications listed in the embodiments describing such uses andmedical indications described herein, are also specifically embraced bythe present invention just as if each and every subcombination of usesand medical indications was individually and explicitly recited herein.

Processes of the Invention

The present invention is directed to, inter alia, processes useful inthe preparation of an anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide, a modulator of thecannabinoid CB₂ receptor.

One aspect of the present invention relates to processes for preparingan anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide comprising thesteps of:

-   -   1) crystallizing        (1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylic        acid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide from a        crystallizing mixture to obtain a crystalline form of        (1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylic        acid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide in the        crystallizing mixture, wherein the crystallizing mixture        comprises acetonitrile and water; and    -   2) isolating the crystalline form of        (1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylic        acid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide from the        crystallizing mixture to obtain the anhydrous crystalline form        of        (1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylic        acid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide.

In some embodiments, crystallizing is conducted at a temperature ofabout −10° C. to about 35° C. In some embodiments, crystallizing isconducted at a temperature of about −10° C. to about 25° C. In someembodiments, crystallizing is conducted at a temperature of about −10°C. to about 10° C. In some embodiments, crystallizing is conducted at atemperature of about −5° C. to about 5° C.

In some embodiments, the crystallizing mixture is prepared by the stepsof:

1) dissolving(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide in acetonitrile anda first amount of water to form a first mixture; and

2) adding a second amount of water to the first mixture to obtain thecrystallizing mixture. In some embodiments, dissolving is conducted at atemperature of about 25° C. to about 80° C. In some embodiments,dissolving is conducted at a temperature of about 40° C. to about 70° C.In some embodiments, dissolving is conducted at a temperature of about55° C. to about 65° C. In some embodiments, dissolving is conducted at atemperature of about 58° C. to about 62° C. In some embodiments,dissolving is conducted at a temperature of about 60° C. In someembodiments, the molar ratio present in the first mixture between(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide, acetonitrile, andfirst amount water is about 1.0:7.3:30.0 to about 1.0:12.1:49.6. In someembodiments, the molar ratio present in the first mixture between(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide, acetonitrile, andfirst amount water is about 1.0:7.8:31.8 to about 1.0:11.6:47.6. In someembodiments, the molar ratio present in the first mixture between(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide, acetonitrile, andfirst amount water is about 1.0:8.2:33.7 to about 1.0:11.2:45.7. In someembodiments, the molar ratio present in the first mixture between(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide, acetonitrile, andfirst amount water is about 1.0:8.7:35.7 to about 1.0:10.7:43.7. In someembodiments, the molar ratio present in the first mixture between(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide, acetonitrile, andfirst amount water is about 1.0:9.2:37.0 to about 1.0:10.2:41.7. In someembodiments, the molar ratio present in the first mixture between(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide, acetonitrile, andfirst amount water is about 1.0:9.7:39.7. In some embodiments, adding ofthe second amount of water to the first mixture is conducted at a ratethat the temperature of the mixture of the second amount of watertogether with the first mixture is at about 25° C. to about 80° C. Insome embodiments, adding of the second amount of water to the firstmixture is conducted at a rate that the temperature of the mixture ofthe second amount of water together with the first mixture is at about40° C. to about 70° C. In some embodiments, adding of the second amountof water to the first mixture is conducted at a rate that thetemperature of the mixture of the second amount of water together withthe first mixture is at about 55° C. to about 65° C. In someembodiments, adding of the second amount of water to the first mixtureis conducted at a rate that the temperature of the mixture of the secondamount of water together with the first mixture is at about 58° C. toabout 62° C. In some embodiments, adding of the second amount of waterto the first mixture is conducted at a rate that the temperature of themixture of the second amount of water together with the first mixture isat about 60° C.

In some embodiments, the molar ratio present in the crystallizingmixture between(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide, acetonitrile, andwater is about 1.0:7.3:327.4 to about 1.0:12.1:545.6. In someembodiments, the molar ratio present in the crystallizing mixturebetween(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide, acetonitrile, andwater in the crystallizing mixture is about 1.0:7.8:349.2 to about1.0:11.6:523.8. In some embodiments, the molar ratio present in thecrystallizing mixture between(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide, acetonitrile, andwater in the crystallizing mixture is about 1.0:8.2:371.0 to about1.0:11.2:502.0. In some embodiments, the molar ratio present in thecrystallizing mixture between(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide, acetonitrile, andwater in the crystallizing mixture is about 1.0:8.7:392.8 to about1.0:10.7:480.1. In some embodiments, the molar ratio present in thecrystallizing mixture between(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide, acetonitrile, andwater in the crystallizing mixture is about 1.0:9.2:414.7 to about1.0:10.2:458.3. In some embodiments, the molar ratio present in thecrystallizing mixture between(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide, acetonitrile, andwater in the crystallizing mixture is about 1.0:9.7:436.5.

In some embodiments,(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide in thecrystallizing and dissolving steps is selected from the group consistingof:

1) a dichloromethane solvate of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide;

2) an acetone solvate of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide;

3) a non-selective solvate of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide; and

4) an ethyl acetate solvate of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide; and

mixtures thereof.

In some embodiments,(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide prior to thedissolving step is selected from the group consisting of:

1) a dichloromethane solvate of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide;

2) an acetone solvate of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide;

3) a non-selective solvate of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide; and

4) an ethyl acetate solvate of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide; and

mixtures thereof.

In some embodiments, isolating comprises filtering the crystalline formof(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide from thecrystallizing mixture.

In some embodiments, isolating comprises removing the crystalline formof(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide from thecrystallizing mixture.

One aspect of the present invention relates to processes for preparingan anhydrous crystalline form, the processes further comprise the stepof drying the crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide to obtain theanhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide. In someembodiments, drying is conducted at a temperature of about 15° C. toabout 80° C. In some embodiments, drying is conducted at a temperatureof about 25° C. to about 65° C. In some embodiments, drying is conductedat a temperature of about 35° C. to about 55° C. In some embodiments,drying is conducted at a temperature of about 50° C. In someembodiments, drying is conducted at a pressure of less than 760 mm Hgand a temperature of about 35° C. to about 55° C. In some embodiments,drying is conducted at a pressure of less than 760 mm Hg and atemperature of about 55° C. to about 65° C.

In some embodiments, after isolating, the anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide has a chemicalpurity of about 95% or greater. In some embodiments, after isolating,the anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide has a chemicalpurity of about 98% or greater. In some embodiments, after isolating,the anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide has a chemicalpurity of about 99% or greater. In some embodiments, after isolating,the anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide has an enantiomericexcess of about 95% or greater. In some embodiments, after isolating,the anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide has an enantiomericexcess of about 98% or greater. In some embodiments, after isolating,the anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide has an enantiomericexcess of about 99% or greater. In some embodiments, after isolating,the anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide has a chemicalpurity of about 99% or greater and an enantiomeric excess of about 99%or greater.

One aspect of the present invention relates to an anhydrous crystallineform of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide prepared by aprocess described herein.

One aspect of the present invention relates to processes of making acomposition comprising mixing an anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide as described hereinwith a pharmaceutically acceptable carrier.

One aspect of the present invention relates to processes of making acomposition further comprising forming the composition into drugproduct, such as, a tablet, a pill, a powder, a lozenge, a sachet, acachet, an elixir, a suspension, an emulsion, a solution, a syrup, asoft gelatin capsule, a hard gelatin capsule, a suppository, a sterileinjectable solution, or a sterile packaged powder.

Crystalline Forms of Compound 1

One aspect of the present invention relates to anhydrous and solvateforms of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1).

One aspect of the present invention relates to DCM solvates of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1).

One aspect of the present invention relates to an anhydrous form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1).

One aspect of the present invention relates to acetone solvates of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-anide (Compound 1).

One aspect of the present invention relates to non-selective solvates of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1).

One aspect of the present invention relates to ethyl acetate solvates of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1).

Crystalline forms of the solvates and anhydrous forms described hereincan be identified by their unique solid state signature with respect to,for example, differential scanning calorimetry (DSC), powder X-raydiffraction (PXRD), and other solid state methods.

Further characterization with respect to water or solvent content ofcrystalline forms can be gauged by any of the following methods forexample, thermogravimetric analysis (TGA), DSC and the like.

For DSC, it is known that the temperatures observed will depend uponsample purity, the rate of temperature change, as well as samplepreparation technique and the particular instrument employed. Thus, thevalues reported herein relating to DSC thermograms can vary by plus orminus about 4° C. (i.e., ±4° C.). The values reported herein relating toDSC thermograms can also vary by plus or minus about 20 joules per gram(i.e., ±20 joules/gram).

In some embodiments, the DSC thermogram values reported herein relate todesolvation events. When DSC thermogram values reported herein relate todesolvation events, the values reported herein are estimates. Scan rateand pan closure can influence DSC values for desolvation events, whichcan vary by plus or minus about 25° C. (i.e., ±25° C.). DSC values fordesolvation events reported herein were recorded using a sample in analuminum pan with an uncrimped lid and a scan rate of 10° C./min.

For PXRD, the relative intensities of the peaks can vary, depending uponthe sample preparation technique, the sample mounting procedure and theparticular instrument employed. Moreover, instrument variation and otherfactors can often affect the 2θ values. Therefore, the peak assignmentsof diffraction patterns can vary by plus or minus 0.2 °2θ (i.e., ±0.2°2θ).

For TGA, the features reported herein can vary by plus or minus about 5°C. (i.e., ±5° C.). The TGA features reported herein can also vary byplus or minus about 2% (i.e., ±2%) weight change due to, for example,sample variation.

Further characterization with respect to hygroscopicity of thecrystalline forms can be gauged by, for example, dynamic moisturesorption (DMS). The DMS features reported herein can vary by plus orminus about 5% (i.e., ±5%) relative humidity. The DMS features reportedherein can also vary by plus or minus about 5% (i.e., ±5%) weightchange.

1. Dichloromethane (DCM) Solvates of Compound 1.

A. Compound 1 (DCM Solvates)

One aspect of the present invention relates to DCM solvates of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1). TheDCM solvates of Compound 1 are characterized by PXRD. The physicalproperties for the DCM solvates as determined by PXRD are summarized inTable 1 below.

TABLE 1 Compound 1 (DCM solvates) PXRD FIG. 2: Peaks of about ≧9.7%relative intensity at 8.3, 9.6, 10.7, 11.0, 15.0, 16.5, 16.7, 17.3, and25.1 °2θ

The amount of DCM present in these solvates can vary, and be up to about10.6% by weight. The amount of DCM can readily be determined by TGA. Thephysical properties for a DCM solvate from Example 1, Method 1, Step Fare summarized in Table 2 below.

TABLE 2 Compound 1 (DCM solvates, Example 1, Method 1, Step F) TGA FIG.1: Decrease in weight of about 5.9% out to about 150° C. DSC FIG. 1:Endotherm extrapolated onset temperature: about 163° C.

Certain powder X-ray diffraction peaks for the DCM solvates of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1) areshown in Table 3 below.

TABLE 3 d-spacing Rel. Int. Pos. [°2θ.] [Å] [%] 6.4 13.8 6.1 8.3 10.6100.0 9.6 9.2 11.3 10.7 8.3 15.6 11.0 8.0 24.3 11.8 7.5 2.2 12.5 7.1 2.713.8 6.4 4.5 14.4 6.1 7.3 15.0 5.9 9.7 15.8 5.6 6.0 16.5 5.4 13.9 16.75.3 30.2 17.3 5.1 16.5 18.2 4.9 2.6 18.7 4.7 5.6 19.5 4.6 2.3 20.4 4.46.1 21.6 4.1 7.7 24.1 3.7 5.3 25.1 3.5 46.0 26.1 3.4 6.3 28.6 3.1 4.329.1 3.1 2.6

B. Dichloromethane Hemi-Solvate of Compound 1

One aspect of the present invention relates to the DCM hemi-solvate of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1). TheDCM hemi-solvate of Compound 1 was prepared by slow crystallization fromCH₂Cl₂ and hexanes (Example 2). The crystal structure was solved and isshown in FIG. 3.

2. Compound 1 (Anhydrous Form).

One aspect of the present invention relates to an anhydrous form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1). Thephysical properties of the crystalline form of Compound 1 (anhydrousform) are summarized in Table 4 below.

TABLE 4 Compound 1 (Anhydrous Form) PXRD FIG. 6: Peaks of about ≧8.7%relative intensity at 8.5, 9.8, 10.7, 11.1, 11.8, 14.5, 16.5, 16.9,17.4, 18.9, 22.1, and 25.4 °2θ TGA FIG. 7: Decrease in weight of about0.24% out to about 150° C. DSC FIG. 7: Endotherm extrapolated onsettemperature: about 162° C. DMS FIG. 7A: The adsorption/desorptionisotherm shows about 1.0% or less weight change from about 10% relativehumidity (RH) to about 90% RH; and about 0.1% or less weight changeafter a 10% RH to 90% RH back to 10% RH cycle, See Example 13.

Certain powder X-ray diffraction peaks for the anhydrous form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1) areshown in Table 5 below.

TABLE 5 d-spacing Pos. [°2θ.] [Å] Rel. Int. [%] 6.6 13.4 4.4 7.9 11.35.5 8.5 10.5 100.0 9.8 9.0 21.5 10.7 8.3 28.3 11.1 7.9 26.1 11.8 7.510.0 13.8 6.4 7.5 14.5 6.1 11.3 14.9 6.0 6.0 16.0 5.6 5.8 16.5 5.4 11.816.9 5.2 26.7 17.3 5.1 8.5 17.4 5.1 14.8 18.4 4.8 4.6 18.9 4.7 8.7 20.24.4 5.5 20.9 4.3 4.4 22.1 4.0 14.5 23.4 3.8 3.0 24.7 3.6 4.6 25.4 3.526.6 26.5 3.4 8.1 29.2 3.1 2.8 29.3 3.0 3.6

One aspect of the present invention relates to an anhydrous crystallineform of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide.

One aspect of the present invention relates to an anhydrous crystallineform of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide, wherein theanhydrous crystalline form has a powder X-ray diffraction patterncomprising a peak, in terms of 2θ, at 8.5°±0.2°. In some embodiments,the anhydrous crystalline form has a powder X-ray diffraction patterncomprising peaks, in terms of 2θ, at 8.5°±0.2°, and 10.7°±0.2°. In someembodiments, the anhydrous crystalline form has a powder X-raydiffraction pattern comprising peaks, in terms of 2θ, at 8.5°±0.2°,10.7°±0.2°, and 16.9°±0.2°. In some embodiments, the anhydrouscrystalline form has a powder X-ray diffraction pattern comprisingpeaks, in terms of 2θ, at 8.5°±0.2°, 10.7°±0.2°, 16.9°±0.2°, 25.4°±0.2°,and 11.1°±0.2°. In some embodiments, the anhydrous crystalline form hasa powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at8.5°±0.2°, 10.7°±0.2°, 16.9°±0.2°, 25.4°±0.2°, 11.1°±0.2°, 9.8°±0.2°,and 17.4°±0.2°. In some embodiments, the anhydrous crystalline form hasa powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at8.5°±0.2°, 10.7°±0.2°, 16.9°±0.2°, 25.4°±0.2°, 11.1°±0.2°, 9.8°±0.2°,17.4°±0.2°, 22.1°±0.2°, and 16.5°±0.2°. In some embodiments, theanhydrous crystalline form has a powder X-ray diffraction patterncomprising peaks, in terms of 2θ, at 8.5°±0.2°, 10.7°±0.2°, 16.9°±0.2°,25.4°±0.2°, 11.1°±0.2°, 9.8°±0.2°, 17.4°±0.2°, 22.1°±0.2°, 16.5°±0.2°,14.5°±0.2°, 11.8°±0.2°, and 18.9°±0.2°. In some embodiments, theanhydrous crystalline form has a powder X-ray diffraction patternsubstantially as shown in FIG. 6, wherein by “substantially” is meantthat the reported peaks can vary by about ±0.2 °2θ.

In some embodiments, the anhydrous crystalline form has a differentialscanning calorimetry thermogram comprising an endotherm with anextrapolated onset temperature between about 159.6° C. and about 169.6°C. In some embodiments, the anhydrous crystalline form has adifferential scanning calorimetry thermogram comprising an endothermwith an extrapolated onset temperature between about 160.6° C. and about168.6° C. In some embodiments, the anhydrous crystalline form has adifferential scanning calorimetry thermogram comprising an endothermwith an extrapolated onset temperature between about 162.6° C. and about166.6° C. In some embodiments, the anhydrous crystalline form has havinga differential scanning calorimetry thermogram comprising an endothermwith an extrapolated onset temperature between about 163.6° C. and about165.6° C. In some embodiments, the anhydrous crystalline form has adifferential scanning calorimetry thermogram comprising an endothermwith an extrapolated onset temperature at about 164.6° C. In someembodiments, the anhydrous crystalline form has a differential scanningcalorimetry thermogram substantially as shown in FIG. 7, wherein by“substantially” is meant that the reported DSC features can vary byabout ±4° C. and that the reported DSC features can vary by about ±20joules per gram.

In some embodiments, the anhydrous crystalline form has athermogravimetric analysis profile showing about 0.5% weight loss belowabout 135° C. In some embodiments, the anhydrous crystalline form has athermogravimetric analysis profile showing about 0.25% weight loss belowabout 135° C. In some embodiments, the anhydrous crystalline form has athermogravimetric analysis profile showing about 0.05% weight loss belowabout 135° C. In some embodiments, the anhydrous crystalline form has athermogravimetric analysis profile substantially as shown in FIG. 7,wherein by “substantially” is meant that the reported TGA features canvary by about ±5° C., and that that the reported TGA features can varyby about ±2% weight change.

One aspect of the present invention relates to the anhydrous crystallineform having:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 8.5°±0.2°, and 10.7°±0.2°;

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature between about 159.6° C.and about 169.6° C.; and/or

3) a thermogravimetric analysis profile showing about 0.5% weight lossbelow about 135° C.

One aspect of the present invention relates to the anhydrous crystallineform having:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 8.5°±0.2°, 10.7°±0.2°, and 16.9°±0.2°;

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature between about 160.6° C.and about 168.6° C.; and/or

3) a thermogravimetric analysis profile showing about 0.25% weight lossbelow about 135° C.

One aspect of the present invention relates to the anhydrous crystallineform having:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 8.5°±0.2°, 10.7°±0.2°, 16.9°±0.2°, 25.4°±0.2°, and 11.1°±0.2°;

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature between about 162.6° C.and about 166.6° C.; and/or

3) a thermogravimetric analysis profile showing about 0.05% weight lossbelow about 135° C.

One aspect of the present invention relates to the anhydrous crystallineform having:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 8.5°±0.2°, 10.7°±0.2°, 16.9°±0.2°, 25.4°±0.2°, 11.1°±0.2°, 9.8°±0.2°,and 17.4°±0.2°;

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature between about 163.6° C.and about 165.6° C.; and/or

3) a thermogravimetric analysis profile showing about 0.05% weight lossbelow about 135° C.

One aspect of the present invention relates to the anhydrous crystallineform having:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 8.5°±0.2°, 10.7°±0.2°, 16.9°±0.2°, 25.4°±0.2°, 11.1°±0.2°, 9.8°±0.2°,17.4°±0.2°, 22.1°±0.2°, and 16.5°±0.2°;

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature at about 164.6° C.;and/or

3) a thermogravimetric analysis profile showing about 0.05% weight lossbelow about 135° C.

One aspect of the present invention relates to the anhydrous crystallineform having:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 8.5°±0.2°, 10.7°±0.2°, 16.9°±0.2°, 25.4°±0.2°, 11.1°±0.2°, 9.8°±0.2°,17.4°±0.2°, 22.1°±0.2°, 16.5°±0.2°, 14.5°±0.2°, 11.8°±0.2°, and18.9°±0.2°;

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature at about 164.6° C.;and/or

3) a thermogravimetric analysis profile showing about 0.05% weight lossbelow about 135° C.

One aspect of the present invention relates to the anhydrous crystallineform having:

1) a powder X-ray diffraction pattern substantially as shown in FIG. 6;

2) a differential scanning calorimetry thermogram substantially as shownin FIG. 7; and/or

3) a thermogravimetric analysis profile substantially as shown in FIG.7.

3. Compound 1 (Acetone Solvates).

One aspect of the present invention relates to acetone solvates of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1). Theacetone solvates of Compound 1 are characterized by PXRD. The physicalproperties for the acetone solvates as determined by PXRD are summarizedin Table 6 below.

TABLE 6 Compound 1 (Acetone Solvate) PXRD FIG. 8: Peaks of about ≧5.4%relative intensity at 7.1, 8.3, 10.1, 11.0, 13.7, 16.1, 16.6, 17.3,22.7, 25.0, 25.6, and 26.0 °2θ

The amount of acetone present in these solvates can vary and can readilybe determined by TGA. The physical properties for a acetone solvate fromExample 5 are summarized in Table 7 below.

TABLE 7 Compound 1 (Acetone Solvate, Example 5) TGA FIG. 9: Decrease inweight of about 5.5% out to about 150° C. DSC FIG. 9: Endothermextrapolated onset temperature: about 163° C.

Certain powder X-ray diffraction peaks for the acetone solvates of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1) areshown in Table 8 below.

TABLE 8 d-spacing Rel. Int. Pos. [°2θ.] [Å] [%] 8.3 10.6266 100.0 25.03.56844 45.0 16.6 5.34216 26.2 17.3 5.1231 23.5 11.0 8.04494 14.4 10.18.74974 9.2 26.0 3.4277 8.8 7.1 12.45547 8.3 22.7 3.91113 7.7 13.76.46525 6.9 16.1 5.51282 6.3 25.6 3.47777 5.4 18.8 4.72926 4.5 15.65.67632 4.4 21.5 4.12408 3.9 6.1 14.41073 3.8 18.1 4.90385 3.5 19.74.51452 3.5 15.5 5.72733 3.3 14.4 6.13613 3.2 24.7 3.61152 3.1 20.84.26245 2.9 9.5 9.35087 2.8 29.8 3.00222 2.8 19.5 4.54403 2.8 19.94.4696 2.6 16.9 5.24284 2.6 11.5 7.71311 2.5 19.1 4.65159 2.4 27.73.22211 2.3

One aspect of the present invention relates to an acetone solvate of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide.

One aspect of the present invention relates to an acetone solvate havinga powder X-ray diffraction pattern comprising a peak, in terms of 2θ, at8.3°±0.2°. In some embodiments, the acetone solvate has a powder X-raydiffraction pattern comprising peaks, in terms of 2θ, at 8.3°±0.2° and25.0°±0.2°. In some embodiments, the acetone solvate has a powder X-raydiffraction pattern comprising peaks, in terms of 2θ, at 8.3°±0.2°,25.0°±0.2°, and 16.6°±0.2°. In some embodiments, the acetone solvate hasa powder X-ray diffraction pattern comprising peaks, in terms of 2θ, at8.3°±0.2°, 25.0°±0.2°, 16.6°±0.2°, 17.3°±0.2°, and 11.0°±0.2°. In someembodiments, the acetone solvate has a powder X-ray diffraction patterncomprising peaks, in terms of 2θ, at 8.3°±0.2°, 25.0°±0.2°, 16.6°±0.2°,17.3°±0.2°, 11.0°±0.2°, 10.1°±0.2°, and 26.0°±0.2°. In some embodiments,the acetone solvate has a powder X-ray diffraction pattern comprisingpeaks, in terms of 2θ, at 8.3°±0.2°, 25.0°±0.2°, 16.6°±0.2°, 17.3°±0.2°,11.0°±0.2°, 10.1°±0.2°, 26.0°±0.2°, 7.1°±0.2°, and 22.7°±0.2°. In someembodiments, the acetone solvate has a powder X-ray diffraction patterncomprising peaks, in terms of 2θ, at 8.3°±0.2°, 25.0°±0.2°, 16.6°±0.2°,17.3°±0.2°, 11.0°±0.2°, 10.1°±0.2°, 26.0°±0.2°, 7.1°±0.2°, 22.7°±0.2°,13.7°±0.2°, 16.1°±0.2°, and 25.6°±0.2°. In some embodiments, the acetonesolvate has a powder X-ray diffraction pattern substantially as shown inFIG. 8, wherein by “substantially” is meant that the reported peaks canvary by about ±0.2°2θ.

In some embodiments, the acetone solvate has a differential scanningcalorimetry thermogram comprising an endotherm with an extrapolatedonset temperature between about 158.0° C. and about 168.0° C. In someembodiments, the acetone solvate has a differential scanning calorimetrythermogram comprising an endotherm with an extrapolated onsettemperature between about 159.0° C. and about 167.0° C. In someembodiments, the acetone solvate has a differential scanning calorimetrythermogram comprising an endotherm with an extrapolated onsettemperature between about 161.0° C. and about 165.0° C. In someembodiments, the acetone solvate has a differential scanning calorimetrythermogram comprising an endotherm with an extrapolated onsettemperature between about 162.0° C. and about 163.0° C. In someembodiments, the acetone solvate has a differential scanning calorimetrythermogram comprising an endotherm with an extrapolated onsettemperature at about 163.0° C. In some embodiments, the acetone solvatehas a differential scanning calorimetry thermogram substantially asshown in FIG. 9, wherein by “substantially” is meant that the reportedDSC features can vary by about ±4° C. and that the reported DSC featurescan vary by about ±20 joules per gram.

In some embodiments, the acetone solvate has a thermnogravimetricanalysis profile showing about 6.0% weight loss below about 150° C. Insome embodiments, the acetone solvate has a thermogravimetric analysisprofile showing about 5.75% weight loss below about 150° C. In someembodiments, the acetone solvate has a thermogravimetric analysisprofile showing about 5.5% weight loss or less below about 150° C. Insome embodiments, the acetone solvate has a thermogravimetric analysisprofile substantially as shown in FIG. 9, wherein by “substantially” ismeant that the reported TGA features can vary by about ±5° C., and thatthat the reported TGA features can vary by about ±2% weight change.

One aspect of the present invention relates to the acetone solvatehaving:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 8.3°±0.2° and 25.0°±0.2°; and/or

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature between about 158.0° C.and about 168.0° C.

One aspect of the present invention relates to the acetone solvatehaving:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 8.3°±0.2°, 25.0°±0.2°, and 16.6°±0.2°; and/or

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature between about 159.0° C.and about 167.0° C.

One aspect of the present invention relates to the acetone solvatehaving:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 8.3°±0.2°, 25.0°±0.2°, 16.6°±0.2°, 17.3°±0.2°, and 11.0°±0.2°; and/or

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature between about 161.0° C.and about 165.0° C.

One aspect of the present invention relates to the acetone solvatehaving:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 8.3°±0.2°, 25.0°±0.2°, 16.6°±0.2°, 17.3°±0.2°, 11.0°±0.2°,10.1°±0.2°, and 26.0°±0.2°;

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature between about 162.0° C.and about 163.0° C.; and/or

3) a thermogravimetric analysis profile showing about 6.0% weight lossor less below about 150° C.

One aspect of the present invention relates to the acetone solvatehaving:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 8.3±0.20, 25.0°±0.2°, 16.6°±0.2°, 17.3°±0.2°, 11.0°±0.2°, 10.1°±0.2°,26.0°±0.2°, 7.1°±0.20, and 22.7°±0.2°;

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature at about 163.0° C.;and/or

3) a thermogravimetric analysis profile showing about 5.75% weight lossbelow about 150° C.

One aspect of the present invention relates to the acetone solvatehaving:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 8.3°±0.2°, 25.0°±0.2°, 16.6°±0.2°, 17.3°±0.2°, 11.0°±0.2°,10.1°±0.2°, 26.0°±0.2°, 7.1°±0.2°, 22.7°±0.2°, 13.7°±0.2°, 16.1°±0.2°,and 25.6°±0.2°;

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature at about 163.0° C.;and/or

3) a thermogravimetric analysis profile showing about 5.5% weight lossor less below about 150° C.

One aspect of the present invention relates to the acetone solvatehaving:

1) a powder X-ray diffraction pattern substantially as shown in FIG. 8;

2) a differential scanning calorimetry thermogram substantially as shownin FIG. 9; and/or

3) a thermogravimetric analysis profile substantially as shown in FIG.9.

4. Compound 1 (Non-Selective Solvates).

One aspect of the present invention relates to non-selective solvates of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1).Non-selective solvates refer to solvates that have substantially thesame crystalline form as determined by PXRD, and depending on thepurity, after de-solvation will have similar extrapolated onsettemperature (+/−4.0° C.) as determined by DSC regardless what solvent orsolvents were used to prepare the solvate. It is understood that the TGAtrace will vary from one non-selective solvate to another and isprimarily determined by the solvent used in the preparation, the solvateformed, and the amount of the solvnet present in the solvate.

The non-selective solvates of Compound 1 are characterized by PXRD. Thephysical properties for the non-selective solvates as determined by PXRDare summarized in Table 9 below.

TABLE 9 Compound 1 (Non-Selective Solvate) PXRD FIG. 10: Peaks of about≧24% relative intensity at 7.9, 9.9, 10.3, 10.7, 10.9, 13.0, 14.9, 16.5,17.4, 18.2, 18.3, 18.9, 19.9, 20.4, 20.5, 21.8, and 23.8 °2θ

The amount of the respective solvent present in these solvates can varyand can readily be determined by TGA. One such non-selective solvate ofCompound 1 is the ethyl acetate solvate as described in Example 6. Thephysical properties (i.e., TGA and DSC) for this ethyl acetatenon-selective solvate are summarized in Table 10 below.

TABLE 10 Compound 1 (Non-Selective Solvate/Ethyl Acetate, Example 6) TGAFIG. 13: Decrease in weight of about 4.8% out to about 150° C. DSC FIG.13: Endotherm extrapolated onset temperature: about 161° C.

Certain powder X-ray diffraction peaks for the non-selective solvates of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1) areshown in Table 11 below.

TABLE 11 d-spacing Pos. [°2θ] [Å] Rel. Int. [%] 7.3 12.14275 16.7 7.911.16727 39.7 8.5 10.44535 11.7 9.9 8.93424 37.9 10.3 8.61737 26.3 10.78.23598 26.3 10.9 8.12357 24.0 11.3 7.81277 10.5 13.0 6.82221 40.8 13.36.64907 9.5 14.9 5.92853 24.0 15.2 5.84616 11.2 16.0 5.53769 21.1 16.25.46327 11.9 16.5 5.35726 24.5 16.9 5.24138 10.7 17.4 5.09682 36.7 18.24.88174 100.0 18.3 4.83505 44.3 18.9 4.70738 24.3 19.2 4.62061 7.0 19.64.53523 13.6 19.9 4.47005 44.2 20.4 4.35851 29.0 20.5 4.33052 32.8 21.24.1998 18.1 21.5 4.13161 14.7 21.8 4.07567 37.1 22.3 3.99444 8.1 23.83.74137 43.5 24.0 3.70405 16.7 24.4 3.64871 12.3 24.7 3.60844 11.6 24.93.57752 20.8 25.1 3.54559 13.0 25.5 3.49243 9.0 26.0 3.43338 8.8 27.73.22622 6.9 27.9 3.19367 7.1 28.3 3.14991 8.6

One aspect of the present invention relates to non-selective solvates of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide.

One aspect of the present invention relates to a non-selective solvatehaving a powder X-ray diffraction pattern comprising a peak, in terms of2θ, at 18.2°±0.2°. In some embodiments, the non-selective solvate has apowder X-ray diffraction pattern comprising peaks, in terms of 2θ, at18.2°±0.2° and 18.3°±0.2°. In some embodiments, the non-selectivesolvate has a powder X-ray diffraction pattern comprising peaks, interms of 2θ, at 18.2°±0.2°, 18.3°±0.2°, and 19.9°±0.2°. In someembodiments, the non-selective solvate has a powder X-ray diffractionpattern comprising peaks, in terms of 2θ, at 18.2°±0.2°, 18.3°±0.2°,19.9°±0.2°, 23.8°±0.2°, and 13.0°±0.2°. In some embodiments, thenon-selective solvate has a powder X-ray diffraction pattern comprisingpeaks, in terms of 2θ, at 18.2°±0.2°, 18.3°±0.2°, 19.9°±0.2°,23.8°±0.2°, 13.0°±0.2°, 7.9°±0.2°, and 9.9°±0.2°. In some embodiments,the non-selective solvate has a powder X-ray diffraction patterncomprising peaks, in terms of 2θ, at 18.2°±0.2°, 18.3°±0.2°, 19.9°±0.2°,23.8°±0.2°, 13.0°±0.2°, 7.9°±0.2°, 9.9°±0.2°, 21.8°±0.2°, and17.4°±0.2°. In some embodiments, the non-selective solvate has a powderX-ray diffraction pattern comprising peaks, in terms of 2θ, at18.2°±0.2°, 18.3°±0.2°, 19.9°±0.2°, 23.8°±0.2°, 13.0°±0.2°, 7.9°±0.2°,9.9°±0.2°, 21.8°±0.2°, 17.4°±0.2°, 20.5°±0.2°, 20.4°±0.2°, 10.7°±0.2°,10.3°±0.2°, 16.5°±0.2°, 18.9°±0.2°, 14.9°±0.2°, and 10.9°±0.2°. In someembodiments, the non-selective solvate has a powder X-ray diffractionpattern substantially as shown in FIG. 10, wherein by “substantially” ismeant that the reported peaks can vary by about ±0.2 °2θ.

In some embodiments, the non-selective solvate has a differentialscanning calorimetry thermogram comprising an endotherm with anextrapolated onset temperature between about 159.8° C. and about 165.8°C. In some embodiments, the non-selective solvate has a differentialscanning calorimetry thermogram comprising an endotherm with anextrapolated onset temperature between about 160.8° C. and about 164.8°C. In some embodiments, the non-selective solvate has a differentialscanning calorimetry thermogram comprising an endotherm with anextrapolated onset temperature between about 158.8° C. and about 162.8°C. In some embodiments, the non-selective solvate has a differentialscanning calorimetry thermogram comprising an endotherm with anextrapolated onset temperature between about 159.8° C. and about 161.8°C. In some embodiments, the non-selective solvate has a differentialscanning calorimetry thermogram comprising an endotherm with anextrapolated onset temperature at about 160.8° C. In some embodiments,the non-selective solvate has a differential scanning calorimetrythermogram substantially as shown in FIG. 13, wherein by “substantially”is meant that the reported DSC features can vary by about ±4° C. andthat the reported DSC features can vary by about ±20 joules per gram.

In some embodiments, the non-selective solvate has a thermogravimetricanalysis profile showing about 5.0% weight loss below about 150° C. Insome embodiments, the non-selective solvate has a thermogravimetricanalysis profile showing about 4.9% weight loss below about 150° C. Insome embodiments, the non-selective solvate has a thermogravimetricanalysis profile showing about 4.8% weight loss below about 150° C. Insome embodiments, the non-selective solvate has a thermogravimetricanalysis profile substantially as shown in FIG. 13, wherein by“substantially” is meant that the reported TGA features can vary byabout ±5° C., and that that the reported TGA features can vary by about±2% weight change.

One aspect of the present invention relates to the non-selective solvatehaving:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 18.2°±0.2° and 18.3°±0.2°; and/or

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature between about 160.8° C.and about 165.8° C.

One aspect of the present invention relates to the non-selective solvatehaving:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 18.2°±0.2°, 18.3°±0.2°, and 19.9°±0.2°; and/or

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature between about 160.8° C.and about 164.8° C.

One aspect of the present invention relates to the non-selective solvatehaving:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 18.2°±0.2°, 18.3°±0.2°, 19.9°±0.2°, 23.8°±0.2°, and 13.0°±0.2°;and/or

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature between about 158.8° C.and about 162.8° C.

One aspect of the present invention relates to the non-selective solvatehaving:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 18.2°±0.2°, 18.3°±0.2°, 19.9°±0.2°, 23.8°±0.2°, 13.0°±0.2°,7.9°±0.2°, and 9.9°±0.2°;

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature between about 159.8° C.and about 161.8° C.; and/or

3) a thermogravimetric analysis profile showing about 5.0% weight lossbelow about 150° C.

One aspect of the present invention relates to the non-selective solvatehaving:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 18.2°±0.20, 18.3°±0.2°, 19.9°±0.2°, 23.8°±0.2°, 13.0°±0.2°,7.9°±0.2°, 9.9°±0.2°, 21.8°±0.2°, and 17.4°±0.2°;

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature at about 160.8° C.;and/or

3) a thermogravimetric analysis profile showing about 4.9% weight lossbelow about 150° C.

One aspect of the present invention relates to the non-selective solvatehaving:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 18.2°±0.2°, 18.3°±0.2°, 19.9°±0.2°, 23.8°±0.2°, 13.0°±0.2°,7.9°±0.2°, 9.9°±0.2°, 21.8°±0.2°, 17.4°±0.2°, 20.5°±0.2°, 20.4°±0.2°,10.7°±0.2°, 10.3°±0.2°, 16.5°±0.2°, 18.9°±0.2°, 14.9°±0.2°, and10.9°±0.2°;

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature at about 160.8° C.;and/or

3) a thermogravimetric analysis profile showing about 4.8% weight lossbelow about 150° C.

One aspect of the present invention relates to the non-selective solvatehaving:

1) a powder X-ray diffraction pattern substantially as shown in FIG. 10;

2) a differential scanning calorimetry thermogram substantially as shownin FIG. 13; and/or

3) a thermogravimetric analysis profile substantially as shown in FIG.13.

5. Compound 1 (Ethyl Acetate Solvate).

One aspect of the present invention relates to the ethyl acetate solvateof(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1). Theethyl acetate solvate of Compound 1 is characterized by PXRD. Thephysical properties for the ethyl acetate solvate as determined by PXRDare summarized in Table 12 below.

TABLE 12 Compound 1 (Ethyl Acetate Solvate) PXRD FIG. 16: Peaks of about≧16.0% relative intensity at 8.1, 8.3, 9.0, 12.8, 14.2, 16.1, 16.7,17.3, 17.9, 18.4, 22.9, 24.7, and 25.7 °2θ

The amount of ethyl acetate present in this solvate can vary but canreadily be determined by TGA. The physical properties for a thenon-selective solvate as the ethyl acetate solvate from Example 7 aresummarized in Table 13 below.

TABLE 13 Compound 1 (Non-Selective Solvate/Ethyl Acetate, Example 7) TGAFIG. 17: Decrease in weight of about 4.7% by weight out to about 120° C.DSC FIG. 17: Endotherm extrapolated onset temperature: about 121° C.

Certain powder X-ray diffraction peaks for the crystalline form of ethylacetate solvates of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1) areshown in Table 14 below.

TABLE 14 d-spacing Rel. Int. Pos. [°2θ.] [Å] [%] 6.0 14.7 9.0 8.1 10.936.8 8.3 10.6 100.0 9.0 9.8 47.5 10.1 8.8 11.6 11.4 7.8 6.4 12.8 6.937.1 14.2 6.3 39.8 14.6 6.1 7.4 15.3 5.8 14.9 16.1 5.5 16.9 16.3 5.413.4 16.7 5.3 37.1 17.3 5.1 24.8 17.9 5.0 27.1 18.4 4.8 27.4 19.7 4.56.7 21.5 4.1 10.1 21.6 4.1 9.8 21.8 4.1 5.7 22.9 3.9 18.6 23.4 3.8 10.723.7 3.8 10.6 24.7 3.6 22.4 25.1 3.5 13.7 25.7 3.5 21.0 26.4 3.4 15.626.8 3.3 4.6 28.5 3.1 6.7 29.4 3.0 5.3

One aspect of the present invention relates to ethyl acetate solvates of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide.

One aspect of the present invention relates to an ethyl acetate solvatehaving a powder X-ray diffraction pattern comprising a peak, in terms of2θ, at 8.3°±0.2°. In some embodiments, the ethyl acetate solvate has apowder X-ray diffraction pattern comprising peaks, in terms of 2θ, at8.3°±0.2° and 9.0°±0.2°. In some embodiments, the ethyl acetate solvatehas a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 8.3°±0.2°, 9.0°±0.2°, and 14.2°±0.2°. In some embodiments, the ethylacetate solvate has a powder X-ray diffraction pattern comprising peaks,in terms of 2θ, at 8.3°±0.2°, 9.0°±0.2°, 14.2°±0.2°, 16.7°±0.2°, and12.8°±0.2°. In some embodiments, the ethyl acetate solvate has a powderX-ray diffraction pattern comprising peaks, in terms of 28, at8.3°±0.2°, 9.0°±0.2°, 14.2°±0.2°, 16.7°±0.2°, 12.8°±0.2°, 8.1°±0.2°, and18.4°±0.2°. In some embodiments, the ethyl acetate solvate has a powderX-ray diffraction pattern comprising peaks, in terms of 28, at8.3°±0.2°, 9.0°±0.2°, 14.2°±0.2°, 16.7°±0.2°, 12.8°±0.2°, 8.1°±0.2°,18.4°±0.2°, 17.9°±0.2°, and 17.3°±0.2°. In some embodiments, the ethylacetate solvate has a powder X-ray diffraction pattern comprising peaks,in terms of 28, at 8.3°±0.2°, 9.0°±0.2°, 14.2°±0.2°, 16.7°±0.2°,12.8°±0.2°, 8.1°±0.2°, 18.4°±0.2°, 17.9°±0.2°, 17.3°±0.2°, 24.7°±0.2°,25.7°±0.2°, 22.9°±0.2°, and 16.1°±0.2°. In some embodiments, the ethylacetate solvate has a powder X-ray diffraction pattern substantially asshown in FIG. 16, wherein by “substantially” is meant that the reportedpeaks can vary by about ±0.2 °2θ.

In some embodiments, the ethyl acetate solvate has a differentialscanning calorimetry thermogram comprising an endotherm with anextrapolated onset temperature between about 116.4° C. and about 126.4°C. In some embodiments, the ethyl acetate solvate has a differentialscanning calorimetry thermogram comprising an endotherm with anextrapolated onset temperature between about 117.4° C. and about 125.4°C. In some embodiments, the ethyl acetate solvate has a differentialscanning calorimetry thermogram comprising an endotherm with anextrapolated onset temperature between about 119.4° C. and about 123.4°C. In some embodiments, the ethyl acetate solvate has a differentialscanning calorimetry thermogram comprising an endotherm with anextrapolated onset temperature between about 120.4° C. and about 122.4°C. In some embodiments, the ethyl acetate solvate has a differentialscanning calorimetry thermogram comprising an endotherm with anextrapolated onset temperature at about 121.4° C. In some embodiments,the ethyl acetate solvate has a differential scanning calorimetrythermogram substantially as shown in FIG. 17, wherein by “substantially”is meant that the reported DSC features can vary by about ±4° C. andthat the reported DSC features can vary by about ±20 joules per gram.

In some embodiments, the ethyl acetate solvate has a thermogravimetricanalysis profile showing about 5.5% weight loss below about 135° C. Insome embodiments, the ethyl acetate solvate has a thermogravimetricanalysis profile showing about 5.4% weight loss below about 135° C. Insome embodiments, the ethyl acetate solvate has a thermogravimetricanalysis profile showing about 5.3% weight loss below about 135° C. Insome embodiments, the ethyl acetate solvate has a thermogravimetricanalysis profile substantially as shown in FIG. 17, wherein by“substantially” is meant that the reported TGA features can vary byabout ±5° C., and that that the reported TGA features can vary by about±2% weight change.

One aspect of the present invention relates to the ethyl acetate solvatehaving:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 8.3°±0.2° and 9.0°±0.2°; and/or

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature between about 116.4° C.and about 126.4° C.

One aspect of the present invention relates to the ethyl acetate solvatehaving:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 8.3°±0.2°, 9.0°±0.2°, and 14.2°±0.2°; and/or

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature between about 117.4° C.and about 125.4° C.

One aspect of the present invention relates to the ethyl acetate solvatehaving:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 8.3°±0.2°, 9.0°±0.2°, 14.2°±0.2°, 16.7°±0.2°, and 12.8°±0.2°; and/or

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature between about 119.4° C.and about 123.4° C.

One aspect of the present invention relates to the ethyl acetate solvatehaving:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 8.3°±0.2°, 9.0°±0.2°, 14.2°±0.2°, 16.7°±0.2°, 12.8°±0.2°, 8.1°±0.2°,and 18.4°±0.2°;

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature between about 120.4° C.and about 122.4° C.; and/or

3) a thermogravimetric analysis profile showing about 5.5% weight lossbelow about 135° C.

One aspect of the present invention relates to the ethyl acetate solvatehaving:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 8.3°±0.2°, 9.0°±0.2°, 14.2°±0.2°, 16.7°±0.2°, 12.8°±0.2°, 8.1°±0.2°,18.4°±0.2°, 17.9°±0.2°, and 17.3°±0.2°;

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature at about 121.4° C.;and/or

3) a thermogravimetric analysis profile showing about 5.4% weight lossbelow about 135° C.

One aspect of the present invention relates to the ethyl acetate solvatehaving:

1) a powder X-ray diffraction pattern comprising peaks, in terms of 2θ,at 8.3°±0.2°, 9.0°±0.2°, 14.2°±0.2°, 16.7°±0.2°, 12.8°±0.2°, 8.1°±0.2°,18.4°±0.2°, 17.9°±0.2°, 17.3°±0.2°, 24.7°±0.2°, 25.7°±0.2°, 22.9°±0.2°,and 16.1°±0.2°;

2) a differential scanning calorimetry thermogram comprising anendotherm with an extrapolated onset temperature at about 121.4° C.;and/or

3) a thermogravimetric analysis profile showing about 5.3% weight lossbelow about 135° C.

One aspect of the present invention relates to the ethyl acetate solvatehaving:

1) a powder X-ray diffraction pattern substantially as shown in FIG. 16;

2) a differential scanning calorimetry thermogram substantially as shownin FIG. 17; and/or

3) a thermogravimetric analysis profile substantially as shown in FIG.17.

The crystalline forms described herein can be prepared by any of thesuitable procedures known in the art for preparing crystallinepolymorphs. In some embodiments the crystalline forms described hereinare prepared according to the Examples. In some embodiments, thecrystalline forms described herein can be prepared by heatingcrystalline forms other than the crystalline forms described herein. Insome embodiments, the crystalline forms described herein can be preparedby recrystallizing crystalline forms other than the crystalline formsdescribed herein.

Compound 1 of the present invention may be prepared according torelevant published literature procedures that are used by one skilled inthe art. Exemplary reagents and procedures for these reactions appearhereinafter in the working Examples. Protection and deprotection may becarried out by procedures generally known in the art (see, for example,Greene, T. W. and Wuts, P. G. M., Protecting Groups in OrganicSynthesis, 3^(rd) Edition, 1999 [Wiley]).

It is understood that the present invention embraces each enantiomer andmixtures thereof. Separation of the individual isomers (such as, bychiral HPLC, recrystallization of diastereoisomeric mixtures and thelike) or selective synthesis (such as, by enantiomeric selectivesyntheses and the like) of the individual isomers is accomplished byapplication of various methods which are well known to practitioners inthe art.

Indications and Methods of Prophylaxis and/or Treatment

In addition to the foregoing beneficial uses for the modulators ofcannabinoid receptor activity disclosed herein, the compounds disclosedherein are useful in the treatment of several additional diseases anddisorders, and in the amelioration of symptoms thereof. Withoutlimitation, these include the following:

1. PAIN

The analgesic properties of cannabinoids have been recognized for manyyears. For example, animal studies have demonstrated that the CB₁/CB₂agonists anandamide, THC, CP55,940 and WIN 55212-2 are effective againstacute and chronic pain from chemical, mechanical, and thermal painstimuli (reviewed in Walker and Huang (2002) Pharmacol. Ther.95:127-135; reviewed in Pacher, P et al. (2006) Pharmacol. Rev. 58(3):389-462). In humans, topical administration of the CB₁/CB₂ agonistHU-210 attenuates capsaicin-induced hyperalgesia and allodynia (Rukwied,R. et al. (2003) Pain 102:283-288), and co-administration of the CB₁/CB₂agonist THC and cannabidiol (nabiximols, trademark Sativex®) providesrelief from cancer-associated pain (GW Pharmaceuticals press releaseJan. 19, 2005, Jun. 19, 2007) and multiple-sclerosis-associated pain andspasticity (GW Pharmaceuticals press release Sep. 27, 2005, Mar. 11,2009).

The role of CB₁ in mediating these analgesic effects is well-documented(reviewed in Manzanares, J. et al. (2006) Current Neuropharmacology4:239-57; reviewed in Pacher, P. et al. (2006) Pharmacol. Rev. 58(3):389-462). For example, blockade of peripheral or central CB₁ leads tohyperalgesia (Richardson, J. D. et al. (1997) Eur. J. Pharmacol.345:145-153; Calignano, A. et al. (1998) Nature 394:277-281), whereasCB₁ activation by exogenous administration of a CB₁ agonistarachidonyl-2-chloroethylamide reduces pain (Furuse, S. et al. (2009)Anesthesiology 111(1):173-86).

Although less well-documented, CB₂ also plays a role in mediatinganalgesic effects of cannabinoids (reviewed in Guindon and IIohmann(2008) Br. J. Pharmacol. 153:319-334). For example, systemic delivery ofthe CB₂-selective agonist AM1241 suppresses hyperalgesia induced in thecarrageenan, capsaicin, and formalin models of inflammatory pain inrodents (reviewed in Guindon and Hohmann (2008) Br. J. Pharmacol.153:319-334). Local (subcutaneous) or systemic administration of AM1241also reverses tactile and thermal hypersensitivity in rats followingligation of spinal nerves in the chronic constriction injury model ofneuropathic pain (Malan, T. P. el al. (2001) Pain 93:239-245; Ibrahim,M. M. et al. (2003) Proc. Natl. Acad. Sci. 100(18):10529-10533), aneffect which is inhibited by treatment with the CB₂-selective antagonistAM630 (Ibrahim, M. M. et al. (2005) Proc. Natl. Acad. Sci.102(8):3093-8). The CB₂-selective agonist GW405833 administeredsystenmically significantly reverses hypersensitivity to mechanicalstimuli in rats following ligation of spinal nerves (Hu, B. et al.(2009) Pain 143:206-212). Thus, CB₂-selective agonists have also beendemonstrated to attenuate pain in experimental models of acute,inflammatory, and neuropathic pain, and hyperalgesia.

Accordingly, CB₂-specific agonists and/or CB₁/CB₂ agonists find use inthe treatment and/or prophylaxis of acute nociception and inflammatoryhyperalgesia, as well as the allodynia and hyperalgesia produced byneuropathic pain. For example, these agonists are useful as an analgesicto treat pain arising from autoimmune conditions; allergic reactions;bone and joint pain; muscle pain; dental pain; nephritic syndrome;scleroderma; thyroiditis; migraine and other headache pain; painassociated with diabetic neuropathy; fibromyalgia, HIV-relatedneuropathy, sciatica, and neuralgias; pain arising from cancer; and painthat occurs as an adverse affect of therapeutics for the treatment ofdisease.

Furthermore, although cannabinoids exert their antinociceptive effectsby complex mechanisms involving effects on the central nervous system,spinal cord, and peripheral sensory nerves (reviewed in Pacher, P. etal. (2006) Pharmacol. Rev. 58(3): 389-462), an analysis of models ofinflammatory and neuropathic pain in mice that are deficient for CB₁only in nociceptive neurons localized in the peripheral nervous systemdemonstrates that the contribution of CB₁-type receptors expressed onthe peripheral terminals of nociceptors to cannabinoid-induced analgesiais paramount (Agarwal, N. et al. (2007) Nat. Neurosci. 10(7): 870-879).Accordingly, CB₁ agonists that are unable to cross the blood brainbarrier still find use in the treatment and/or prophylaxis of acutepain, inflanunatory pain, neuropathic pain, and hyperalgesia.

2. DISORDERS OF THE IMMUNE SYSTEM

Autoimmune Disorders.

Cannabinoid receptor agonists have been demonstrated to attenuateaberrant immune responses in autoimmune disorders, and in some cases, toprovide protection to the tissue that is being inappropriately targetedby the immune system.

For example, Multiple Sclerosis (MS) is an autoimmune disorder thatresults in the demyelination of neurons in the CNS. The CB₁/CB₂ agonistTHC significantly inhibits the severity of clinical disease in theExperimental Autoimmune Encephalomyelitis (EAE) mouse model of MS, aneffect that is believed to be mediated by CB₁ on neurons and CB₂ onimmune cell (Maresz, K. et al. (2007) Nat. Med. 13(4):492-497).Consistent with these results, CB₁-selective agonist WIN 55212-2provides significant neuroprotection in the experimental allergicuveitis (EAU) model in mice (Pryce, G. et al. (2003) Brain126:2191-2202), whereas CB₂-selective agonist HU-308 markedly reducesthe recruitment of immature myeloid cells and T cells, microglial andinfiltrating myeloid cell proliferation, and axonal loss in the EAEmodel (Palazuelos, J. et al. (2008). J. Biol. Chem. 283(19):13320-9).Likewise, the CB₁/CB₂ agonist WIN 55212-2 significantly inhibitsleukocyte rolling and adhesion in the brain in the EAE mouse model, aneffect that is blocked by the CB₂-selective antagonist SR144528 but notthe CB₁-selective antagonist SR141716A (Ni, X. et al. Mult. Sclerosis10(2):158-64). Accordingly, CB₂-selective agonists and/or CB₁/CB₂agonists find use in the treatment and/or prophylaxis of MultipleSclerosis and related autoimmune demyelinating diseases, e.g.Guillan-Barré syndrome, polyradiculoneuropathy and chronic inflammatorydemyelination.

As another example, the autoimmune disease Rheumatoid Arthritis (RA) isa chronic, systemic inflammatory disorder of the skeletal system thatprincipally attacks the joints to produce an inflammatory synovitis andthat often progresses to destruction of the articular cartilage andankylosis of the joints. The CB₁/CB₂ agonists WIN 55212-2 and HU-210significantly inhibit IL-1 alpha-stimulated proteoglycan and collagendegradation in bovine nasal cartilage explants in vitro (Mbvundula, E.et al. (2006) J. Pharm. and Pharmacol. 58:351-358). Accordingly,CB₂-selective agonists and/or CB₁/CB₂ agonists find use in the treatmentand/or prophylaxis of autoimmune arthritic diseases, for example,rheumatoid arthritis, psoriatic arthritis, ankylosing spondylarthritis,and reactive arthritis.

Type 1 Hypersensitivity and Allergic Response.

Cannabinoid receptor agonists have been demonstrated to attenuateaberrant immune responses in allergic reactions as well. In type-1, orimmediate, hypersensitivity, plasma cells that have been activated by anallergen secrete IgE antibodies, which bind to Jc receptors on thesurface of tissue mast cells and blood basophils and eosinophils.Repeated exposure to the same allergen results in cross-linking of thebound IgE on sensitized cells, resulting in secretion ofpharmacologically active mediators such as histamine, leukotriene andprostaglandin. These mediators are responsible for the symptomsassociated with allergies, including vasodilation and increasedpermeability, smooth muscle spasms, and leukocyte extravasation. Topicaladministration of the CB₁/CB₂ agonist HU-210 reduces thesehistamine-induced responses in human skin (Dvorak, M. et al. (2003)Inflamm. Res. 52:238-245). Similarly, subcutaneous injection of CB₁/CB₂agonist THC or increased levels of endogenous cannabinoids reducescutaneous inflammation and the pruritis (itch) associated with it in amouse model for allergic contact dermatitis. (Karsak et al. (2007)Science, 316(5830), 1494-1497). In contrast, injection of the CB₁receptor antagonist S141716A or the CB₂ receptor antagonist SR144528exacerbates this inflammation and pruritis. (Karsak et al. (2007)Science, 316(5830), 1494-1497). Accordingly, CB₂-selective agonistsand/or CB₁/CB₂ agonists find use in the treatment of allergic reactionsincluding atopic dermatitis (pruritis/itch), urticaria (hives), asthma,conjunctivitis, allergic rhinitis (hay fever), and anaphylaxis.

Conditions Associated with CNS Inflammation.

CB₂ agonists have been demonstrated to attenuate inflammation in theCNS. For example, administration of CB₂ agonists prevents the activationof microglia in rodent models of Alzheimer's Disease (Ashton J. C., etal. (2007) Curr. Neuropharmacol. 5(2):73-80). Likewise, administrationof CB₂ agonists reduces the volume of infarcts by 30% in a rodentocclusion model of stroke (Zhang, M. et al. (2007) J. Cereb. Blood FlowMetab. 27:1387-96). Thus, CB₂ agonists find use in the treatment and/orprophylaxis of neuropathologies associated with CNS inflammation, e.g.Alzheimer's, stroke-induced damage, dementia, ALS, and HIV.

Conditions Associated with Vascular Inflammation.

CB₂ is expressed in macrophages and T cells in atherosclerotic plaques,and the CB₁/CB₂ agonist THC reduces the progression of atherosclerosisin ApoE knockout mice, a well studied mouse model of atherosclerosis.The CB₂-specific antagonist SR144528 completely blocks this effect invitro and in vivo (Steffens, S. et al. (2005) Nature 434:782-786). Thus,CB₂ agonists find use in treating atherosclerosis.

Other Disorders Associated with Aberrant or Unwanted Immune Response.

Given the expression of CB₂ on a number of different types of immunecells and the attenuating effects that CB₂ agonists have been observedto have on the activities of these cells, CB₂ agonists are useful forthe treatment and/or prophylaxis of other disorders wherein undesiredimmune cell activity and/or inflammation is observed. Such exemplarydisorders include osteoarthritis, anaphylaxis, Behcet's disease, graftrejection, vasculitis, gout, spondylitis, viral and bacterial diseases,e.g. AIDS, and meningitis; and other autoimmune disorders such as lupus,e.g. systemic lupus erythematosus; inflammatory bowel disease, e.g.Crohn's disease, ulcerative colitis; psoriasis; autoimmune hepatitis;and type 1 diabetes mellitus.

3. BONE AND JOINT DISEASES

Osteoporosis.

CB₂ is expressed in osteoblasts, osteocytes, and osteoclasts.Osteoblasts make new bone, whereas osteoclasts degrade it. TheCB₂-specific agonist HU-308 enhances endocortical osteoblast numbers andactivity while simultaneously inhibiting proliferation of osteoclastprecursors in bone marrow-derived osteoblasts/stromal cells in vitro,and attenuates ovariectomy-induced bone loss and stimulates corticalthickness by stimulating endocortical bone formation and suppressingosteoclast number in vivo (Ofek, O. et al. (2006) Proc. Natl. Acad. Sci.103(3):696-701). Thus, CB₂ agonists are useful for the treatment and/orprophylaxis of disease wherein bone density is decreased, such asosteoporosis.

Arthritis.

As discussed above, CB₂-selective agonists and CB₁/CB₂ agonists areuseful for the treatment and/or prophylaxis of autoimmune arthriticdiseases, for example, rheumatoid arthritis, psoriatic arthritis,ankylosing spondylarthritis, and reactive arthritis, and for thetreatment and/or prophylaxis of inflammation associated withosteoarthritis. In addition, as discussed above, CB₁-selective agonistsand CB₁/CB₂ agonists are useful for the treatment of pain associatedwith these arthritic disorders.

4. EYE DISEASE

Retinal pigment epithelial (RPE) cells provide trophic support tophotoreceptor cells in the eye, and RPE cell death has been demonstratedto be a major contributor to Age-related Macular Degeneration (AMD). TheCB₁/CB₂ agonist CP55940 significantly protects RPE cells from oxidativedamage; the CB₂ receptor agonist, JWH015 provides comparable protection(Wei, Y. et al. (2009) Mol. Vis. 15:1243-51). Accordingly, CB₂-selectiveagonists find use in preventing the onset or progression of vision lossassociated with AMD.

5. COUGH

The cough reflex is predominantly under the control of two classes ofsensory afferent nerve fibers, the myelinated A-delta fibers and thenon-myelinated C-fibers, the activation of which (i.e. depolarization)elicits cough via the vagus nerve afferent pathway. The CB₁/CB₂ agonistCP55940 reduces capsaicin-, PGE₂- and hypertonic saline-induceddepolarization of guinea pig and human vagus nerve preparations in vitro(Patel, H. J. et al. (2003) British J. Pharma. 140:261-8). The CB₁/CB₂agonists WIN 55212-2 produced a dose-dependent inhibition of the numberof capsaicin-induced coughs in mice (Morita, K. et al. (2003) Eur. J.Pharmacol. 474:269-272). The CB₁/CB₂ agonist anandamide produced adose-dependent inhibition of the number of capsaicin-induced coughs inguinea pigs (Calignano, A. et al. (2000) Nature 408:96-101).CB₁-specific antagonist SR141716A attenuates the antitussive effects ofWN 55212-2 and anandamide (Morita, K. et al. (2003) Eur. J. Pharmacol.474:269-272; Calignano, A. et atl. (2000) Nature 408:96-101). TheCB₂-selective agonist JWH133 reduces capsaicin-, PGE₂- and hypertonicsaline-induced depolarization of guinea pig and human vagus nervepreparations in vitro, and administration of CB₂-selective agonistJWH133 prior to exposure to the tussive agent citric acid significantlyreduces cough in conscious guinea-pigs (Patel, H. J. et al. (2003)British J. Pharma. 140:261-8). Thus, both CB₁ and CB₂ play an importantrole in mediating the antitussive effect of cannabinoids, andCB₁-selective agonists and CB₁/CB₂ agonists are useful in the treatmentand/or prophylaxis of cough.

6. CANCER

A number of human leukemia and lymphoma cell lines, including Jurkat,Molt-4 and Sup-T1, express CB₂ and not CB₁, and agonists of CB₂ induceapoptosis in these and primary acute lymphoblastic leukemia (ALL) cells(Nagarkatti, L. C. et al. US2004/0259936). Similarly, CB₂ is expressedon glioblastoma cell lines and treatment with agonists of CB₂ inducesapoptosis of these cells in vitro (Widmer, M. (2008) J. Neurosci. Res.86(14):3212-20). Accordingly, CB₂-selective agonists are useful inattenuating the growth of a malignancy of the immune system, forexample, leukemias, lymphomas, and solid tumors of the glial lineage.

In addition, as discussed above, CB₁-selective agonists and CB₁/CB₂agonists are useful in providing relief from pain associated with cancer(GW Pharmaceuticals press release Jan. 19, 2005, Jun. 19, 2007).

CB₂-mediated signaling is involved in the in vivo and in vitro growthinhibition of prostate cancer cells, which suggests that CB₂ agonistshave potential therapeutic interest in the management of prostatecancer. (Inhibition of human tumour prostate PC-3 cell growth bycannabinoids R(+)-Methanandamide and JWH-015: Involvement of CB₂;Olea-Herrero, et al. British Journal of Cancer advance onlinepublication 18 Aug. 2009; doi: 10.1038/sj.bjc. 6605248).

7. REGENERATIVE MEDICINE

Agonists of CB₂ modulate the expansion of the progenitor pool of neuronsin the CNS. CB₂ antagonists inhibit the proliferation of cultured neuralstem cells and the proliferation of progenitor cells in the SVZ of younganimals, whereas CB₂-selective agonists stimulate progenitor cellproliferation in vivo, with this effect being more pronounced in olderanimals (Goncalves, M. B. et al. (2008) Mol. Cell Neurosci.38(4):526-36). Thus, agonists of CB₂ are useful in regenerativemedicine, for example to promote the expansion of progenitor cells forthe replacement of neurons lost during injury or disease, such asAlzheimer's Disease, stroke-induced damage, dementia, amyotrophiclateral sclerosis (ALS) and Parkinson's Disease.

8. CERTAIN EMBODIMENTS

One aspect of the present invention relates to methods for the treatmentof a cannabinoid receptor-mediated disorder in an individual, comprisingadministering to the individual in need thereof, a therapeuticallyeffective amount of an anhydrous crystalline form as described herein ora pharmaceutical composition thereof.

One aspect of the present invention relates to methods for the treatmentof a CB₂ receptor-mediated disorder in an individual, comprisingadministering to the individual in need thereof, a therapeuticallyeffective amount of an anhydrous crystalline form as described herein ora pharmaceutical composition thereof.

One aspect of the present invention relates to methods for the treatmentof pain in an individual, comprising administering to the individual inneed thereof, a therapeutically effective amount of an anhydrouscrystalline form as described herein or a pharmaceutical compositionthereof.

One aspect of the present invention relates to methods for the treatmentof bone pain in an individual, comprising administering to theindividual in need thereof, a therapeutically effective amount of ananhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof joint pain in an individual, comprising administering to theindividual in need thereof, a therapeutically effective amount of ananhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof pain associated with osteoarthritis in an individual, comprisingadministering to the individual in need thereof, a therapeuticallyeffective amount of an anhydrous crystalline form as described herein ora pharmaceutical composition thereof.

One aspect of the present invention relates to methods for the treatmentof osteoarthritis in an individual, comprising administering to theindividual in need thereof, a therapeutically effective amount of ananhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof osteoporosis in an individual, comprising administering to theindividual in need thereof, a therapeutically effective amount of ananhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof hyperalgesia in an individual, comprising administering to theindividual in need thereof, a therapeutically effective amount of ananhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof allodynia in an individual, comprising administering to theindividual in need thereof, a therapeutically effective amount of ananhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof inflammatory pain in an individual, comprising administering to theindividual in need thereof, a therapeutically effective amount of ananhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof inflammatory hyperalgesia in an individual, comprising administeringto the individual in need thereof, a therapeutically effective amount ofan anhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof neuropathic pain in an individual, comprising administering to theindividual in need thereof, a therapeutically effective amount of ananhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof neuropathic hyperalgesia in an individual, comprising administeringto the individual in need thereof, a therapeutically effective amount ofan anhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof acute nociception in an individual, comprising administering to theindividual in need thereof, a therapeutically effective amount of ananhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof muscle pain in an individual, comprising administering to theindividual in need thereof, a therapeutically effective amount of ananhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof dental pain in an individual, comprising administering to theindividual in need thereof, a therapeutically effective amount of ananhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof migraine and other headache pain in an individual, comprisingadministering to the individual in need thereof, a therapeuticallyeffective amount of an anhydrous crystalline form as described herein ora pharmaceutical composition thereof.

One aspect of the present invention relates to methods for the treatmentof pain that occurs as an adverse effect of therapeutics in anindividual, comprising administering to the individual in need thereof,a therapeutically effective amount of an anhydrous crystalline form asdescribed herein or a pharmaceutical composition thereof.

One aspect of the present invention relates to methods for the treatmentof pain associated with a disorder selected from: cancer, multiplesclerosis, allergic reactions, nephritic syndrome, scleroderma,thyroiditis, diabetic neuropathy, fibromyalgia, HIV related-neuropathy,sciatica, and autoimmune conditions, in an individual, comprisingadministering to the individual in need thereof, a therapeuticallyeffective amount of an anhydrous crystalline form as described herein ora pharmaceutical composition thereof.

One aspect of the present invention relates to methods for the treatmentof multiple sclerosis-associated spasticity in an individual, comprisingadministering to the individual in need thereof, a therapeuticallyeffective amount of an anhydrous crystalline form as described herein ora pharmaceutical composition thereof.

One aspect of the present invention relates to methods for the treatmentof autoimmune disorders in an individual, comprising administering tothe individual in need thereof, a therapeutically effective amount of ananhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof an autoimmune disorder selected from the group consisting of:multiple sclerosis, Guillan-Barré syndrome, polyradiculoneuropathy,chronic inflammatory demyelination, rheumatoid arthritis, psoriaticarthritis, ankylosing spondylarthritis, and reactive arthritis, in anindividual, comprising administering to the individual in need thereof,a therapeutically effective amount of an anhydrous crystalline form asdescribed herein or a pharmaceutical composition thereof.

One aspect of the present invention relates to methods for the treatmentof allergic reactions in an individual, comprising administering to theindividual in need thereof, a therapeutically effective amount of ananhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof an allergic reaction associated with a disorder selected from: atopicdermatitis, pruritis, urticaria, asthma, conjunctivitis, allergicrhinitis, and anaphylaxis in an individual, comprising administering tothe individual in need thereof, a therapeutically effective amount of ananhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof CNS inflammation in an individual, comprising administering to theindividual in need thereof, a therapeutically effective amount of ananhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof CNS inflammation associated with a disorder selected from:Alzheimer's disease, stroke, dementia, amyotrophic lateral sclerosis,and human immnunodeficiency virus, in an individual, comprisingadministering to the individual in need thereof, a therapeuticallyeffective amount of an anhydrous crystalline form as described herein ora pharmaceutical composition thereof.

One aspect of the present invention relates to methods for the treatmentof atherosclerosis in an individual, comprising administering to theindividual in need thereof, a therapeutically effective amount of ananhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof undesired immune cell activity and inflammation associated with adisorder selected from: osteoarthritis, anaphylaxis, Behcet's disease,graft rejection, vasculitis, gout, spondylitis, viral disease, bacterialdisease, lupus, inflammatory bowel disease, autoimmune hepatitis, andtype 1 diabetes mellitus, in an individual, comprising administering tothe individual in need thereof, a therapeutically effective amount of ananhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof age-related macular degeneration in an individual, comprisingadministering to the individual in need thereof, a therapeuticallyeffective amount of an anhydrous crystalline form as described herein ora pharmaceutical composition thereof.

One aspect of the present invention relates to methods for the treatmentof cough in an individual, comprising administering to the individual inneed thereof, a therapeutically effective amount of an anhydrouscrystalline form as described herein or a pharmaceutical compositionthereof.

One aspect of the present invention relates to methods for the treatmentof leukemia in an individual, comprising administering to the individualin need thereof, a therapeutically effective amount of an anhydrouscrystalline form as described herein or a pharmaceutical compositionthereof.

One aspect of the present invention relates to methods for the treatmentof lymphoma in an individual, comprising administering to the individualin need thereof, a therapeutically effective amount of an anhydrouscrystalline form as described herein or a pharmaceutical compositionthereof.

One aspect of the present invention relates to methods for the treatmentof CNS tumors in an individual, comprising administering to theindividual in need thereof, a therapeutically effective amount of ananhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof prostate cancer in an individual, comprising administering to theindividual in need thereof, a therapeutically effective amount of ananhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof Alzheimer's disease in an individual, comprising administering to theindividual in need thereof, a therapeutically effective amount of ananhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof stroke-induced damage in an individual, comprising administering tothe individual in need thereof, a therapeutically effective amount of ananhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to methods for the treatmentof dementia in an individual, comprising administering to the individualin need thereof, a therapeutically effective amount of an anhydrouscrystalline form as described herein or a pharmaceutical compositionthereof.

One aspect of the present invention relates to methods for the treatmentof amyotrophic lateral sclerosis in an individual, comprisingadministering to the individual in need thereof, a therapeuticallyeffective amount of an anhydrous crystalline form as described herein ora pharmaceutical composition thereof.

One aspect of the present invention relates to methods for the treatmentof Parkinson's disease in an individual, comprising administering to theindividual in need thereof, a therapeutically effective amount of ananhydrous crystalline form as described herein or a pharmaceuticalcomposition thereof.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of a cannabinoid receptor-mediated disorder.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of a CR₂ receptor-mediated disorder.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of pain.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of bone pain.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of joint pain.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of pain associated with osteoarthritis.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of osteoarthritis.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of osteoporosis.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of hyperalgesia.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of allodynia.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of inflammatory pain.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of inflammatory hyperalgesia.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of neuropathic pain.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of neuropathic hyperalgesia.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of acute nociception.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of muscle pain.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of dental pain.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of migraine and other headache pain.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of pain that occurs as an adverse effect oftherapeutics.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of pain associated with a disorder selected from:cancer, multiple sclerosis, allergic reactions, nephritic syndrome,scleroderma, thyroiditis, diabetic neuropathy, fibromyalgia, HIVrelated-neuropathy, sciatica, and autoimmune conditions.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of multiple sclerosis-associated spasticity.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of autoimmune disorders.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of an autoimmune disorder selected from the groupconsisting of: multiple sclerosis, Guillan-Barré syndrome,polyradiculoneuropathy, chronic inflammatory demyelination, rheumatoidarthritis, psoriatic arthritis, ankylosing spondylarthritis, andreactive arthritis.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of allergic reactions.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of an allergic reaction associated with a disorderselected from: atopic dermatitis, pruritis, urticaria, asthma,conjunctivitis, allergic rhinitis, and anaphylaxis.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of CNS inflammation.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of CNS inflammation associated with a disorderselected from: Alzheimer's disease, stroke, dementia, amyotrophiclateral sclerosis, and human immunodeficiency virus.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of atherosclerosis.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of undesired immune cell activity and inflammationassociated with a disorder selected from: osteoarthritis, anaphylaxis,Behcet's disease, graft rejection, vasculitis, gout, spondylitis, viraldisease, bacterial disease, lupus, inflammatory bowel disease,autoimmune hepatitis, and type 1 diabetes mellitus.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of age-related macular degeneration.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of cough.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of leukemia.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of lymphoma.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of CNS tumors.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of prostate cancer.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of Alzheimer's disease.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of stroke-induced damage.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of dementia.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of amyotrophic lateral sclerosis.

One aspect of the present invention relates to the use of an anhydrouscrystalline form as described herein, in the manufacture of a medicamentfor the treatment of Parkinson's disease.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of the humanor animal body by therapy.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of acannabinoid receptor-mediated disorder.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of a CB₂receptor-mediated disorder.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of pain.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of bone pain.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of jointpain.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of painassociated with osteoarthritis.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment ofosteoarthritis.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment ofosteoporosis.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment ofhyperalgesia.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of allodynia.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment ofinflammatory pain.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment ofinflammatory hyperalgesia.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment ofneuropathic pain.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment ofneuropathic hyperalgesia.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of acutenociception.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of musclepain.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of dentalpain.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of migraineand other headache pain.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of pain thatoccurs as an adverse effect of therapeutics.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of painassociated with a disorder selected from: cancer, multiple sclerosis,allergic reactions, nephritic syndrome, scleroderma, thyroiditis,diabetic neuropathy, fibromyalgia, HIV related-neuropathy, sciatica, andautoimmune conditions.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of multiplesclerosis-associated spasticity.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of autoimmunedisorders.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of anautoimmune disorder selected from the group consisting of: multiplesclerosis, Guillan-Barré syndrome, polyradiculoneuropathy, chronicinflammatory demyelination, rheumatoid arthritis, psoriatic arthritis,ankylosing spondylarthritis, and reactive arthritis.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of allergicreactions.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of anallergic reaction associated with a disorder selected from: atopicdermatitis, pruritis, urticaria, asthma, conjunctivitis, allergicrhinitis, and anaphylaxis.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of CNSinflammation.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of CNSinflammation associated with a disorder selected from: Alzheimer'sdisease, stroke, dementia, amyotrophic lateral sclerosis, and humanimmunodeficiency virus.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment ofatherosclerosis.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of undesiredimmune cell activity and inflammation associated with a disorderselected from: osteoarthritis, anaphylaxis, Behcet's disease, graftrejection, vasculitis, gout, spondylitis, viral disease, bacterialdisease, lupus, inflammatory bowel disease, autoimmune hepatitis, andtype 1 diabetes mellitus.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment ofage-related macular degeneration.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of cough.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of leukemia.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of lymphoma.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of CNStumors.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of prostatecancer.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment ofAlzheimer's disease.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment ofstroke-induced damage.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment of dementia.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment ofamyotrophic lateral sclerosis.

One aspect of the present invention relates to an anhydrous crystallineform as described herein, for use in a method of treatment ofParkinson's disease.

Pharmaceutical Compositions and Dosage Forms

One aspect of the present invention relates to compositions comprisingan anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide as describedherein.

One aspect of the present invention relates to compositions comprisingan anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide as describedherein, and a pharmaceutically acceptable carrier.

One aspect of the present invention relates to pharmaceuticalcompositions comprising an anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide as describedherein, and a pharmaceutically acceptable carrier.

One aspect of the present invention relates to dosage forms comprisingan anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide as describedherein, and a pharmaceutically acceptable carrier.

One aspect of the present invention relates to processes for preparingpharmaceutical compositions comprising the steps of:

1) preparing an anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide according to any ofthe processes described herein; and

2) admixing said anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide with apharmaceutically acceptable carrier.

One aspect of the present invention relates to processes for preparing adosage form comprising the steps of:

1) preparing an anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide according to any ofthe processes described herein; and

2) admixing said anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide with apharmaceutically acceptable carrier.

One aspect of the present invention relates to compositions comprising asolvate as described herein.

Some embodiments of the present invention include a method of producinga pharmaceutical composition comprising admixing at least one compoundaccording to any of the compound embodiments disclosed herein and apharmaceutically acceptable carrier.

Formulations may be prepared by any suitable method, typically byuniformly mixing the active compound(s) with liquids or finely dividedsolid carriers, or both, in the required proportions and then, ifnecessary, forming the resulting mixture into a desired shape ordispensing into a desired vial or ampule.

Conventional excipients, such as binding agents, fillers, acceptablewetting agents, tabletting lubricants and disintegrants may be used intablets and capsules for oral administration. Liquid preparations fororal administration may be in the form of solutions, emulsions, aqueousor oily suspensions and syrups. Alternatively, the oral preparations maybe in the form of dry powder that can be reconstituted with water oranother suitable liquid vehicle before use. Additional additives such assuspending or emulsifying agents, non-aqueous vehicles (including edibleoils), preservatives and flavorings and colorants may be added to theliquid preparations. Parenteral dosage forms may be prepared bydissolving the compound of the invention in a suitable liquid vehicleand filter sterilizing the solution before filling and sealing anappropriate vial or ampule. These are just a few examples of the manyappropriate methods well known in the art for preparing dosage forms.

A compound of the present invention can be formulated intopharmaceutical compositions using techniques well known to those in theart. Suitable pharmaceutically-acceptable carriers, outside thosementioned herein, are known in the art; for example, see Remington, TheScience and Practice of Pharmacy, 20^(th) Edition, 2000, LippincottWilliams & Wilkins, (Editors: Gennaro et al.)

While it is possible that, for use in the prophylaxis or treatment, acompound of the invention may, in an alternative use, be administered asa raw or pure chemical, it is preferable however to present the compoundor active ingredient as a pharmaceutical formulation or compositionfurther comprising a pharmaceutically acceptable carrier.

Pharmaceutical formulations include those suitable for oral, rectal,nasal, topical (including buccal and sub-lingual), vaginal or parenteral(including intramuscular, subcutaneous and intravenous) administrationor in a form suitable for administration by inhalation, insufflation orby a transdermal patch. Transdermal patches dispense a drug at acontrolled rate by presenting the drug for absorption in an efficientmanner with minimal degradation of the drug. Typically, transdermalpatches comprise an impermeable backing layer, a single pressuresensitive adhesive and a removable protective layer with a releaseliner. One of ordinary skill in the art will understand and appreciatethe techniques appropriate for manufacturing a desired efficacioustransdermal patch based upon the needs of the artisan.

The compounds of the invention, together with a conventional adjuvant,carrier, or diluent, may thus be placed into the form of pharmaceuticalformulations and unit dosage forms thereof and in such form may beemployed as solids, such as tablets or filled capsules, or liquids suchas solutions, suspensions, emulsions, elixirs, gels or capsules filledwith the same, all for oral use, in the form of suppositories for rectaladministration; or in the form of sterile injectable solutions forparenteral (including subcutaneous) use. Such pharmaceuticalcompositions and unit dosage forms thereof may comprise conventionalingredients in conventional proportions, with or without additionalactive compounds or principles and such unit dosage forms may containany suitable effective amount of the active ingredient commensurate withthe intended daily dosage range to be employed.

For oral administration, the pharmaceutical composition may be in theform of, for example, a tablet, capsule, suspension or liquid. Thepharmaceutical composition is preferably made in the form of a dosageunit containing a particular amount of the active ingredient. Examplesof such dosage units are capsules, tablets, powders, granules or asuspension, with conventional additives such as lactose, mannitol, cornstarch or potato starch; with binders such as crystalline cellulose,cellulose derivatives, acacia, corn starch or gelatins; withdisintegrators such as corn starch, potato starch or sodiumcarboxymethyl-cellulose; and with lubricants such as talc or magnesiumstearate. The active ingredient may also be administered by injection asa composition wherein, for example, saline, dextrose or water may beused as a suitable pharmaceutically acceptable carrier.

Compounds of the present invention or a solvate, hydrate orphysiologically functional derivative thereof can be used as activeingredients in pharmaceutical compositions, specifically as cannabinoidreceptor modulators. By the term “active ingredient” is defined in thecontext of a “pharmaceutical composition” and refers to a component of apharmaceutical composition that provides the primary pharmacologicaleffect, as opposed to an “inactive ingredient” which would generally berecognized as providing no pharmaceutical benefit.

The dose when using the compounds of the present invention can varywithin wide limits and as is customary and is known to the physician, itis to be tailored to the individual conditions in each individual case.It depends, for example, on the nature and severity of the illness to betreated, on the condition of the patient, on the compound employed or onwhether an acute or chronic disease state is treated or prophylaxisconducted or on whether further active compounds are administered inaddition to the compounds of the present invention. Representative dosesof the present invention include, but not limited to, about 0.001 mg toabout 5000 mg, about 0.001 mg to about 2500 mg, about 0.001 mg to about1000 mg, 0.001 mg to about 500 mg, 0.001 mg to about 250 mg, about 0.001mg to 100 mg, about 0.001 mg to about 50 mg and about 0.001 mg to about25 mg. Multiple doses may be administered during the day, especiallywhen relatively large amounts are deemed to be needed, for example 2, 3or 4 doses. Depending on the individual and as deemed appropriate fromthe patient's physician or caregiver it may be necessary to deviateupward or downward from the doses described herein.

The amount of active ingredient, or an active salt or derivativethereof, required for use in treatment will vary not only with theparticular salt selected but also with the route of administration, thenature of the condition being treated and the age and condition of thepatient and will ultimately be at the discretion of the attendantphysician or clinician. In general, one skilled in the art understandshow to extrapolate in vivo data obtained in a model system, typically ananimal model, to another, such as a human. In some circumstances, theseextrapolations may merely be based on the weight of the animal model incomparison to another, such as a mammal, preferably a human, however,more often, these extrapolations are not simply based on weights, butrather incorporate a variety of factors. Representative factors includethe type, age, weight, sex, diet and medical condition of the patient,the severity of the disease, the route of administration,pharmacological considerations such as the activity, efficacy,pharmacokinetic and toxicology profiles of the particular compoundemployed, whether a drug delivery system is utilized, on whether anacute or chronic disease state is being treated or prophylaxis conductedor on whether further active compounds are administered in addition tothe compounds of the present invention and as part of a drugcombination. The dosage regimen for treating a disease condition withthe compounds and/or compositions of this invention is selected inaccordance with a variety factors as cited above. Thus, the actualdosage regimen employed may vary widely and therefore may deviate from apreferred dosage regimen and one skilled in the art will recognize thatdosage and dosage regimen outside these typical ranges can be testedand, where appropriate, may be used in the methods of this invention.

The desired dose may conveniently be presented in a single dose or asdivided doses administered at appropriate intervals, for example, astwo, three, four or more sub-doses per day. The sub-dose itself may befurther divided, e.g., into a number of discrete loosely spacedadministrations. The daily dose can be divided, especially whenrelatively large amounts are administered as deemed appropriate, intoseveral, for example 2, 3 or 4 part administrations. If appropriate,depending on individual behavior, it may be necessary to deviate upwardor downward from the daily dose indicated.

The compounds of the present invention can be administrated in a widevariety of oral and parenteral dosage forms. It will be obvious to thoseskilled in the art that the following dosage forms may comprise, as theactive component, either a compound of the invention or apharmaceutically acceptable salt, solvate or hydrate of a compound ofthe invention.

For preparing pharmaceutical compositions from the compounds of thepresent invention, the selection of a suitable pharmaceuticallyacceptable carrier can be either solid, liquid or a mixture of both.Solid form preparations include powders, tablets, pills, capsules,cachets, suppositories and dispersible granules. A solid carrier can beone or more substances which may also act as diluents, flavoring agents,solubilizers, lubricants, suspending agents, binders, preservatives,tablet disintegrating agents, or an encapsulating material.

In powders, the carrier is a finely divided solid which is in a mixturewith the finely divided active component.

In tablets, the active component is mixed with the carrier having thenecessary binding capacity in suitable proportions and compacted to thedesire shape and size.

The powders and tablets may contain varying percentage amounts of theactive compound. A representative amount in a powder or tablet maycontain from 0.5 to about 90 percent of the active compound; however, anartisan would know when amounts outside of this range are necessary.Suitable carriers for powders and tablets are magnesium carbonate,magnesium stearate, talc, sugar, lactose, pectin, dextrin, starch,gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, alow melting wax, cocoa butter and the like. The term “preparation”includes the formulation of the active compound with encapsulatingmaterial as carrier providing a capsule in which the active component,with or without carriers, is surrounded by a carrier, which is thus inassociation with it. Similarly, cachets and lozenges are included.Tablets, powders, capsules, pills, cachets and lozenges can be used assolid forms suitable for oral administration.

For preparing suppositories, a low melting wax, such as an admixture offatty acid glycerides or cocoa butter, is first melted and the activecomponent is dispersed homogeneously therein, as by stirring. The moltenhomogenous mixture is then poured into convenient sized molds, allowedto cool and thereby to solidify.

Formulations suitable for vaginal administration may be presented aspessaries, tampons, creams, gels, pastes, foams or sprays containing inaddition to the active ingredient such carriers as are known in the artto be appropriate.

Liquid form preparations include solutions, suspensions and emulsions,for example, water or water-propylene glycol solutions. For example,parenteral injection liquid preparations can be formulated as solutionsin aqueous polyethylene glycol solution. Injectable preparations, forexample, sterile injectable aqueous or oleaginous suspensions may beformulated according to the known art using suitable dispersing orwetting agents and suspending agents. The sterile injectable preparationmay also be a sterile injectable solution or suspension in a nontoxicparenterally acceptable diluent or solvent, for example, as a solutionin 1,3-butanediol. Among the acceptable vehicles and solvents that maybe employed are water, Ringer's solution and isotonic sodium chloridesolution. In addition, sterile, fixed oils are conventionally employedas a solvent or suspending medium. For this purpose any bland fixed oilmay be employed including synthetic mono- or diglycerides. In addition,fatty acids such as oleic acid find use in the preparation ofinjectables.

The compounds according to the present invention may thus be formulatedfor parenteral administration (e.g. by injection, for example bolusinjection or continuous infusion) and may be presented in unit dosageform in ampoules, pre-filled syringes, small volume infusion or inmulti-dose containers with an added preservative. The pharmaceuticalcompositions may take such forms as suspensions, solutions, or emulsionsin oily or aqueous vehicles and may contain formulatory agents such assuspending, stabilizing and/or dispersing agents. Alternatively, theactive ingredient may be in powder form, obtained by aseptic isolationof sterile solid or by lyophilization from solution, for constitutionwith a suitable vehicle, e.g. sterile, pyrogen-free water, before use.

Aqueous formulations suitable for oral use can be prepared by dissolvingor suspending the active component in water and adding suitablecolorants, flavors, stabilizing and thickening agents, as desired.

Aqueous suspensions suitable for oral use can be made by dispersing thefinely divided active component in water with viscous material, such asnatural or synthetic gums, resins, methylcellulose, sodiumcarboxymethylcellulose, or other well-known suspending agents.

Also included are solid form preparations which can be converted,shortly before use, to liquid form preparations for oral administration.Such liquid forms include solutions, suspensions and emulsions. Thesepreparations may contain, in addition to the active component,colorants, flavors, stabilizers, buffers, artificial and naturalsweeteners, dispersants, thickeners, solubilizing agents and the like.

For topical administration to the epidermis the compounds according tothe invention may be formulated as ointments, creams or lotions, or as atransdermal patch.

Ointments and creams may, for example, be formulated with an aqueous oroily base with the addition of suitable thickening and/or gellingagents. Lotions may be formulated with an aqueous or oily base and willin general also contain one or more emulsifying agents, stabilizingagents, dispersing agents, suspending agents, thickening agents, orcoloring agents.

Formulations suitable for topical administration in the mouth includelozenges comprising active agent in a flavored base, usually sucrose andacacia or tragacanth; pastilles comprising the active ingredient in aninert base such as gelatin and glycerin or sucrose and acacia; andmouthwashes comprising the active ingredient in a suitable liquidcarrier.

Solutions or suspensions are applied directly to the nasal cavity byconventional means, for example with a dropper, pipette or spray. Theformulations may be provided in single or multi-dose form. In the lattercase of a dropper or pipette, this may be achieved by the patientadministering an appropriate, predetermined volume of the solution orsuspension. In the case of a spray, this may be achieved for example bymeans of a metering atomizing spray pump.

Administration to the respiratory tract may also be achieved by means ofan aerosol formulation in which the active ingredient is provided in apressurized pack with a suitable propellant. If the compounds of thepresent invention or pharmaceutical compositions comprising them areadministered as aerosols, for example as nasal aerosols or byinhalation, this can be carried out, for example, using a spray, anebulizer, a pump nebulizer, an inhalation apparatus, a metered inhaleror a dry powder inhaler. Pharmaceutical forms for administration of thecompounds of the present invention as an aerosol can be prepared byprocesses well known to the person skilled in the art. For theirpreparation, for example, solutions or dispersions of the compounds ofthe present invention in water, water/alcohol mixtures or suitablesaline solutions can be employed using customary additives, for examplebenzyl alcohol or other suitable preservatives, absorption enhancers forincreasing the bioavailability, solubilizers, dispersants and othersand, if appropriate, customary propellants, for example include carbondioxide, CFCs, such as, dichlorodifluoromethane, trichlorofluoromethane,or dichlorotetrafluoroethane; and the like. The aerosol may convenientlyalso contain a surfactant such as lecithin. The dose of drug may becontrolled by provision of a metered valve.

In formulations for administration to the respiratory tract, includingintranasal formulations, the compound will generally have a smallparticle size for example of the order of 10 microns or less. Such aparticle size may be obtained by means known in the art, for example bymicronization. When desired, formulations adapted to give sustainedrelease of the active ingredient may be employed.

Alternatively the active ingredients may be provided in the form of adry powder, for example, a powder mix of the compound in a suitablepowder base such as lactose, starch, starch derivatives such ashydroxypropylmethyl cellulose and polyvinylpyrrolidone (PVP).Conveniently the powder carrier will form a gel in the nasal cavity. Thepowder composition may be presented in unit dosage form for example incapsules or cartridges of, e.g., gelatin, or blister packs from whichthe powder may be administered by means of an inhaler.

The pharmaceutical preparations are preferably in unit dosage forms. Insuch form, the preparation is subdivided into unit doses containingappropriate quantities of the active component. The unit dosage form canbe a packaged preparation, the package containing discrete quantities ofpreparation, such as packeted tablets, capsules and powders in vials orampoules. Also, the unit dosage form can be a capsule, tablet, cachet,or lozenge itself, or it can be the appropriate number of any of thesein packaged form.

Tablets or capsules for oral administration and liquids for intravenousadministration are preferred compositions.

The compounds according to the invention may optionally exist aspharmaceutically acceptable salts including pharmaceutically acceptableacid addition salts prepared from pharmaceutically acceptable non-toxicacids including inorganic and organic acids. Representative acidsinclude, but are not limited to, acetic, benzenesulfonic, benzoic,camphorsulfonic, citric, ethenesulfonic, dichloroacetic, formic,fumaric, gluconic, glutamic, hippuric, hydrobromic, hydrochloric,isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic,nitric, oxalic, pamoic, pantothenic, phosphoric, succinic, sulfiric,tartaric, oxalic, p-toluenesulfonic and the like. Certain compounds ofthe present invention which contain a carboxylic acid functional groupmay optionally exist as pharmaceutically acceptable salts containingnon-toxic, pharmaceutically acceptable metal cations and cations derivedfrom organic bases. Representative metals include, but are not limitedto, aluminium, calcium, lithium, magnesium, potassium, sodium, zinc andthe like. In some embodiments the pharmaceutically acceptable metal issodium. Representative organic bases include, but are not limited to,benzathine (N¹,N²-dibenzylethane-1,2-diamine), chloroprocaine(2-(diethylamino)ethyl 4-(chloroamino)benzoate), choline,diethanolamine, ethylenediamine, meglumine((2R,3R,4R,5S)-6-(methylamino)hexane-1,2,3,4,5-pentaol), procaine(2-(diethylalmino)ethyl 4-aminobenzoate), and the like. Certainpharmaceutically acceptable salts are listed in Berge, et al., Journalof Pharmaceutical Sciences, 66:1-19 (1977).

The acid addition salts may be obtained as the direct products ofcompound synthesis. In the alternative, the free base may be dissolvedin a suitable solvent containing the appropriate acid and the saltisolated by evaporating the solvent or otherwise separating the salt andsolvent. The compounds of this invention may form solvates with standardlow molecular weight solvents using methods known to the skilledartisan.

Compounds of the present invention can be converted to “pro-drugs.” Theterm “pro-drugs” refers to compounds that have been modified withspecific chemical groups known in the art and when administered into anindividual these groups undergo biotransformation to give the parentcompound. Pro-drugs can thus be viewed as compounds of the inventioncontaining one or more specialized non-toxic protective groups used in atransient manner to alter or to eliminate a property of the compound. Inone general aspect, the “pro-drug” approach is utilized to facilitateoral absorption. A thorough discussion is provided in T. Higuchi and V.Stella, Pro-drugs as Novel Delivery Systems Vol. 14 of the A.C.S.Symposium Series; and in Bioreversible Carriers in Drug Design, ed.Edward B. Roche, American Pharmaceutical Association and Pergamon Press,1987, both of which are hereby incorporated by reference in theirentirety.

Some embodiments of the present invention include a method of producinga pharmaceutical composition for “combination-therapy” comprisingadmixing at least one compound according to any of the compoundembodiments disclosed herein, together with at least one knownpharmaceutical agent as described herein and a pharmaceuticallyacceptable carrier.

It is noted that when the cannabinoid receptor modulators are utilizedas active ingredients in a pharmaceutical composition, these are notintended for use only in humans, but in other non-human mammals as well.Indeed, recent advances in the area of animal health-care mandate thatconsideration be given for the use of active agents, such as cannabinoidreceptor modulators, for the treatment of a cannabinoidreceptor-associated disease or disorder in companionship animals (e.g.,cats, dogs, etc.) and in livestock animals (e.g., cows, chickens, etc.)Those of ordinary skill in the art are readily credited withunderstanding the utility of such compounds in such settings.

Hydrates and Solvates

It is understood that when the phrase “pharmaceutically acceptablesalts, solvates, and hydrates” or the phrase “pharmaceuticallyacceptable salt, solvate, or hydrate” is used when referring tocompounds described herein, it embraces pharmaceutically acceptablesolvates and/or hydrates of the compounds, pharmaceutically acceptablesalts of the compounds, as well as pharmaceutically acceptable solvatesand/or hydrates of pharmaceutically acceptable salts of the compounds.It is also understood that when the phrase “pharmaceutically acceptablesolvates and hydrates” or the phrase “pharmaceutically acceptablesolvate or hydrate” is used when referring to salts described herein, itembraces pharmaceutically acceptable solvates and/or hydrates of suchsalts.

It will be apparent to those skilled in the art that the dosage formsdescribed herein may comprise, as the active component, either acompound described herein or a pharmaceutically acceptable salt or as apharmaceutically acceptable solvate or hydrate thereof. Moreover,various hydrates and solvates of the compounds described herein andtheir salts will find use as intermediates in the manufacture ofpharmaceutical compositions. Typical procedures for making andidentifying suitable hydrates and solvates, outside those mentionedherein, are well known to those in the art; see for example, pages202-209 of K. J. Guillory, “Generation of Polymorphs, Hydrates,Solvates, and Amorphous Solids,” in: Polymorphism in PharmaceuticalSolids, ed. Harry G. Britain, Vol. 95, Marcel Dekker, Inc., New York,1999. Accordingly, one aspect of the present invention pertains tomethods of administering hydrates and solvates of compounds describedherein and/or their pharmaceutical acceptable salts, that can beisolated and characterized by methods known in the art, such as,thermogravimetric analysis (TGA), TGA-mass spectroscopy, TGA-Infraredspectroscopy, powder X-ray diffraction (XRPD), Karl Fisher titration,high resolution X-ray diffraction, and the like. There are severalcommercial entities that provide quick and efficient services foridentifying solvates and hydrates on a routine basis. Example companiesoffering these services include Wilmington PharmaTech (Wilmington,Del.), Avantium Technologies (Amsterdam) and Aptuit (Greenwich, Conn.).

Polymorphs and Pseudopolymorphs

Polymorphism is the ability of a substance to exist as two or morecrystalline phases that have different arrangements and/or conformationsof the molecules in the crystal lattice. Polymorphs show the sameproperties in the liquid or gaseous state but they behave differently inthe solid state.

Besides single-component polymorphs, drugs can also exist as salts andother multicomponent crystalline phases. For example, solvates andhydrates may contain an API host and either solvent or water molecules,respectively, as guests. Analogously, when the guest compound is a solidat room temperature, the resulting form is often called a cocrystal.Salts, solvates, hydrates, and cocrystals may show polymorphism as well.Crystalline phases that share the same API host, but differ with respectto their guests, may be referred to as pseudopolymorphs of one another.

Solvates contain molecules of the solvent of crystallization in adefinite crystal lattice. Solvates, in which the solvent ofcrystallization is water, are termed hydrates. Because water is aconstituent of the atmosphere, hydrates of drugs may be formed rathereasily.

By way of example, Stahly recently published a polymorph screens of 245compounds consisting of a “wide variety of structural types” revealedthat about 90% of them exhibited multiple solid forms. Overall,approximately half the compounds were polymorphic, often having one tothree forms. About one-third of the compounds formed hydrates, and aboutone-third formed solvates. Data from cocrystal screens of 64 compoundsshowed that 60% formed cocrystals other than hydrates or solvates. (G.P. Stahly, Crystal Growth & Design (2007), 7(6), 1007-1026).

Other Utilities

Another object of the present invention relates to radio-labeledcompounds of the present invention that would be useful not only inradio-imaging but also in assays, both in vitro and in vivo, forlocalizing and quantitating cannabinoid receptors in tissue samples,including human and for identifying cannabinoid receptor ligands byinhibition binding of a radio-labeled compound. It is a further objectof this invention to develop novel cannabinoid receptor assays of whichcomprise such radio-labeled compounds.

The present invention embraces isotopically-labeled crystalline forms ofthe present invention. Isotopically or radio-labeled compounds are thosewhich are identical to compounds disclosed herein, but for the fact thatone or more atoms are replaced or substituted by an atom having anatomic mass or mass number different from the atomic mass or mass numbermost commonly found in nature. Suitable radionuclides that may beincorporated in compounds of the present invention include but are notlimited to ²H (also written as D for deuterium), ³H (also written as Tfor tritium), ¹¹C, ¹³C, ¹⁴C, ¹³N, ¹⁵N, ¹⁵O, ¹⁷O, ¹⁸O, ¹⁸F, ³⁵S, ³⁶Cl,⁷⁵Br, ⁷⁶Br, ⁷⁷Br, ⁸²Br, ¹²³I, ¹²⁴I, ¹²⁵I and ¹³¹I. The radionuclide thatis incorporated in the instant radio-labeled compounds will depend onthe specific application of that radio-labeled compound. For example,for in vitro cannabinoid receptor labeling and competition assays,compounds that incorporate ³H, ¹⁴C, ⁸²Br, ¹²⁵I, ¹³¹I or ³⁵S willgenerally be most useful. For radio-imaging applications ¹¹C, ¹⁸F, ¹²I,¹²³I, ¹²⁴I, ¹³¹I, ⁷⁵Br, ⁷⁶Br or ⁷⁷Br will generally be most useful.

It is understood that a “radio-labeled” or “labeled compound” is acrystalline form of Compound 1 that has incorporated at least oneradionuclide; in some embodiments the radionuclide is selected from thegroup consisting of ³H, and ¹⁴C.

Certain isotopically-labeled crystalline forms of the present inventionare useful in compound and/or substrate tissue distribution assays. Insome embodiments the radionuclide ³H and/or ¹⁴C isotopes are useful inthese studies. Further, substitution with heavier isotopes such asdeuterium (i.e., ²H) may afford certain therapeutic advantages resultingfrom greater metabolic stability (e.g., increased in vivo half-life orreduced dosage requirements) and hence may be preferred in somecircumstances. Isotopically labeled crystalline forms of the presentinvention can generally be prepared by following procedures analogous tothose disclosed in the and Examples infra, by substituting anisotopically labeled reagent for a non-isotopically labeled reagent.Other synthetic methods that are useful are discussed infra. Moreover,it should be understood that all of the atoms represented in thecompounds of the invention can be either the most commonly occurringisotope of such atoms or the scarcer radio-isotope or nonradioactiveisotope.

Synthetic methods for incorporating radio-isotopes into organiccompounds are applicable to compounds of the invention and are wellknown in the art. These synthetic methods, for example, incorporatingactivity levels of tritium into target molecules, are as follows:

A. Catalytic Reduction with Tritium Gas: This procedure normally yieldshigh specific activity products and requires halogenated or unsaturatedprecursors.

B. Reduction with Sodium Borohydride [³H]: This procedure is ratherinexpensive and requires precursors containing reducible functionalgroups such as aldehydes, ketones, lactones, esters and the like.

C. Reduction with Lithium Aluminum Hydride [³H]: This procedure offersproducts at almost theoretical specific activities. It also requiresprecursors containing reducible functional groups such as aldehydes,ketones, lactones, esters and the like.

D. Tritium Gas Exposure Labeling: This procedure involves exposingprecursors containing exchangeable protons to tritium gas in thepresence of a suitable catalyst.

E. N-Methylation using Methyl Iodide [³H]: This procedure is usuallyemployed to prepare O-methyl or N-methyl (3H) products by treatingappropriate precursors with high specific activity methyl iodide (3H).This method in general allows for higher specific activity, such as forexample, about 70-90 Ci/mmol.

Synthetic methods for incorporating activity levels of ¹²⁵I into targetmolecules include:

A. Sandmeyer and like reactions: This procedure transforms an aryl amineor a heteroaryl amine into a diazonium salt, such as a diazoniumtetrafluoroborate salt and subsequently to ¹²⁵I labeled compound usingNa¹²⁵I. A represented procedure was reported by Zhu, G-D. and co-workersin J. Org. Chem., 2002, 67, 943-948.

B. Ortho ¹²⁵Iodination of phenols: This procedure allows for theincorporation of ¹²⁵I at the ortho position of a phenol as reported byCollier, T. L. and co-workers in J. Labelled Compd. Radiopharm., 1999,42, S264-S266.

C. Aryl and heteroaryl bromide exchange with ¹²⁵I: This method isgenerally a two step process. The first step is the conversion of thearyl or heteroaryl bromide to the corresponding tri-alkyltinintermediate using for example, a Pd catalyzed reaction [i.e. Pd(Ph₃P)₄]or through an aryl or heteroaryl lithium, in the presence of atri-alkyltinhalide or hexaalkylditin [e.g., (CH₃)₃SnSn(CH₃)₃]. Arepresentative procedure was reported by Le Bas, M.-D. and co-workers inJ. Labelled Compd. Radiopharm. 2001, 44, S280-S282.

A radiolabeled cannabinoid receptor compound can be used in a screeningassay to identify/evaluate compounds. In general terms, a newlysynthesized or identified compound (i.e., test compound) can beevaluated for its ability to reduce binding of the “radio-labeledCompound 1” to a cannabinoid receptor. Accordingly, the ability of atest compound to compete with the “radio-labeled Compound 1” for thebinding to a cannabinoid receptor directly correlates to its bindingaffinity.

Certain labeled compounds of the present invention bind to certaincannabinoid receptors. In one embodiment the labeled compound has anIC₅₀ less than about 500 μM, in another embodiment the labeled compoundhas an IC₅₀ less than about 100 μM, in yet another embodiment thelabeled compound has an IC₅₀ less than about 10 μM, in yet anotherembodiment the labeled compound has an IC₅₀ less than about 1 μM and instill yet another embodiment the labeled inhibitor has an IC₅₀ less thanabout 0.1 μM.

Other uses of the disclosed receptors and methods will become apparentto those skilled in the art based upon, inter alia, a review of thisdisclosure.

As will be recognized, the steps of the methods of the present inventionneed not be performed any particular number of times or in anyparticular sequence. Additional objects, advantages and novel featuresof this invention will become apparent to those skilled in the art uponexamination of the following examples thereof, which are illustrativeand not limiting.

EXAMPLES Example 1: Preparation of(1aS,5aS)-2-(4-Oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicAcid ((S)-1-Hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1)

Step A: Preparation of (1S,5R)-bicyclo[3.1.0]hexan-2-one

A 2.5 M hexane solution of n-BuLi (489 mL, 1223 mmol) was added dropwiseto a stirred solution of (S)-2-(but-3-enyl)oxirane (100 g, 1019 mmol)and 2,2,6,6-tetramethylpiperidine (86 mL, 509 mmol) in MTBE (1000 mL)cooled in a dry ice/acetone bath, at a rate to maintain the internaltemperature at −12 to −5° C. (time of addition=1 h). After addition wascomplete, the reaction was stirred another hour at −5 to 0° C.

While still at 0° C., 3 M aqueous HCl (545 mL) was added (dropwise atfirst) with stirring (internal temperature rose to 3° C.). The layerswere separated and the organic layer washed with another 200 mL 3 M HCl.The combined aqueous washings were extracted with MTBE (2×500 mL). Thecombined organic layers were washed with brine (3×300 mL) thenconcentrated (at 350 mbar and 29° C. water bath) to ca 1000 mL of paleyellow solution. This solution was carried on without furtherpurification.

To 407 mL water was added dibasic potassium phosphate (216 g, 1240mmol), monobasic potassium phosphate (12.8 g, 94 mmol), and potassiumbromide (18.19 g, 153 mmol). pH paper indicated a pH of ˜9. This aqueoussolution was added to the MTBE solution of(1S,2S,5R)-bicyclo[3.1.0]hexan-2-ol in a 5 L 3-neck round bottom flaskequipped with an overhead stirrer. The mixture was cooled to −20° C. ina dry-ice/isopropanol bath. TEMPO (4.30 g, 27.5 mmol) was added. Thetemperature was allowed to warm to 0° C. and aqueous 10-13% sodiumhypochlorite (1059 mL, 1630 mmol) was added dropwise while maintainingthe internal temperature between −10 and 0° C. (time of addition=70min). Stirring was continued at 0° C. for another hour. 50 g sodiumsulfite was added to quench excess sodium hypochlorite (temperature roseto 12° C.). The layers were separated and the aqueous layer wasextracted twice more with MTBE (500 mL then 250 mL). The combinedorganic layers (total volume ca 1600 mL) were dried (MgSO₄) thenfiltered. The solution was concentrated to ca 300 mL at 300 mbar and 35°C. water bath. The product was distilled—first with house vacuum and a50° C. water bath which distilled off most of the remaining MTBE. Thevacuum pump was connected giving a vacuum of ˜2 torr and the productdistilled (2 torr/36° C.) to give the title compound (65.8 g) as a lightorange oil (note: receiving flask was cooled in dry ice/acetone bath).¹H NMR (400 MHz, CDCl₃) δ 0.93 (td, J=4.6, 3.3 Hz, 1H), 1.20 (td, J=8.0,4.8 Hz, 1H), 1.74-1.79 (m, 1H), 1.98-2.19 (m, 5H).

Step B: Preparation of 2-Hydrazinylpyrazine

The reaction was run under nitrogen atmosphere. 2-chloropyrazine (96 mL,1073 mmol) was added dropwise to 35 wt % aqueous hydrazine (544 mL, 6009mmol) at 65° C. over 1 h. After the addition, stirring was continued at63-67° C. for 16 h then let stand at room temperature for two days. Themixture was filtered to remove a small amount of precipitate, thenextracted with 10% iPrOH/dichloromethane (5×250 mL). The combinedorganic extracts were dried (MgSO₄), filtered, then concentrated underreduced pressure. The resulting solid was triturated with isopropylacetate (600 mL). The solid was collected by filtration, rinsed withisopropyl acetate and dried under vacuum to give 2-hydrazinylpyrazine(60 g, 51%) as a pale yellow solid. LCMS m/z=111.2 [M+H]⁺. ¹H NMR (400MHz, DMSO-d₆) δ 4.21 (s, 2H), 7.70 (d, J=2.8 Hz, 1H), 7.89 (s, 1H), 7.93(dd, J=2.8, 1.5 Hz, 1H), 8.10 (d, J=1.5 Hz, 1H).

Step C: Preparation of(1aS,5aS)-2-(Pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicAcid Ethyl Ester

To a solution of (1S,5R)-bicyclo[3.1.0]hexan-2-one (52.9 g, 539 mmol)and diethyl oxalate (0.073 L, 539 mmol) in absolute ethanol (0.9 L) (notdenatured with methanol) was added a 1.0 M THF solution of potassiumtert-butoxide (0.539 L, 539 mmol) over 15 min (maintaining thetemperature below 43° C.). The resulting yellow solution was stirred at40° C. for 3.5 h (a precipitate appeared within 10 min and the reactioneventually became a thick suspension). 2-hydrazinylpyrazine (59.4 g, 539mmol) was added followed by a 6.0 M aqueous solution of hydrogenchloride (0.270 L, 1618 mmol). The reaction was stirred at 50° C. for1.5 h. The mixture was poured into ice-water (5 L). A precipitateappeared immediately. After standing for 30 minutes in an ice bath, thesolid was collected by filtration, rinsed with water (5×1 L), thendried, affording the title product (106 g, 73%) as an off-white solid.LCMS m/z=271.2 (M+H⁺). ¹H NMR (400 MHz, CDCl₃) δ 0.47 (td, J=4.7, 3.3Hz, 1H), 1.27 (td, J=8.0, 4.9 Hz, 1H), 1.41 (t, J=7.1 Hz, 3H), 2.26-2.32(m, 1H), 2.77-2.82 (m, 1H), 2.88 (dd, J=16.7, 1.4 Hz, 1H), 2.99 (dd,J=16.6, 6.4 Hz, 1H), 4.40 (q, J=7.1 Hz, 2H), 8.41 (dd, J=2.5, 1.5 Hz,1H), 8.52 (d, J=2.5 Hz, 1H), 9.40 (d, J=1.5 Hz, 1H).

Step D: Preparation of(1aS,5aS)-2-(Pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicAcid

To a suspension of(1aS,5aS)-2-(pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ethyl ester (106 g, 392 mmol) in MeOH (300 mL) and THF (300 mL) wasadded a 2.0 M aqueous solution of NaOH (235 mL, 471 mmol). The mixturewas stirred at 23° C. for 20 h. The organic solvents were removed bydistillation. The remaining aqueous solution was diluted with water toca 1.5 L then acidified to pH ˜2 with 6 M HCl (ca 95 mL). The resultingfine precipitate was collected by filtration, rinsed with water, thendried, to give the title compound (95 g, 100%) as a white solid. LCMSm/z=243.1 (M+H⁺). ¹H NMR (400 MHz, DMSO-d6) δ 0.43 (td, J=4.6, 3.2 Hz,1H), 1.26 (td, J=8.0, 4.4 Hz, 1H), 2.27-2.33 (m, 1H), 2.71-2.75 (m, 1H),2.76 (d, J=16.8 Hz, 1H), 2.89 (dd, J=16.4, 6.4 Hz, 1H), 8.61 (dd, J=2.7,1.5 Hz, 1H), 8.67 (d, J=2.5 Hz, 1H), 9.17 (d, J=1.5 Hz, 1H), 13.02 (s,1H).

Two separate methods were used to prepare Compound 1, one methodoxidized the pyrazinyl ring nitrogen as the last reaction step while thesecond method oxidized the pyrazinyl ring nitrogen of the carboxylicacid intermediate (i.e., title compound in of Step D) prior to thecoupling with (S)-2-amino-3,3-dimethylbutan-1-ol. Steps E and F areshown below.

Method 1 Step E: Preparation of(1aS,5aS)-2-Pyrazin-2-yl-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide

To a solution of(1aS,5aS)-2-(pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid (1.4 g, 5.78 mmol) and triethylamine (1.611 mL, 11.56 mmol) in DMF(15 mL) was added HATU (2.242 g, 5.90 mmol). The reaction was stirred at23° C. for 5 min, then was added (S)-2-amino-3,3-dimethylbutan-1-ol(0.711 g, 6.07 mmol). The reaction was stirred at 23° C. for 15 min thenconcentrated. The residue was purified by silica gel flashchromatography (35 to 100% EtOAc/hexanes) to give the title product(1.97 g, 100%) as a white solid. LCMS m/z=342.2 [M+H]+. ¹H NMR (400 MHz,CDCl₃) δ 0.48 (td, J=4.6, 3.4 Hz, 1H), 1.05 (s, 9H), 1.24 (td, J=8.0,4.7 Hz, 1H), 2.26-2.32 (m, 1H), 2.74-2.78 (m, 1H), 2.94 (d, J=16.8 Hz,1H), 3.01 (dd, J=16.7, 6.1 Hz, 1H), 3.67-3.72 (m, 1H), 3.93-3.98 (m,2H), 7.08 (d, J=8.5 Hz, 1H), 8.42 (dd, J=1.4, 0.9 Hz, 1H), 8.51 (d,J=2.7 Hz, 1H), 9.26 (d, J=1.1 Hz, 1H).

Step F: Preparation of(1aS,5aS)-2-(4-Oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1)

To a solution of(1aS,5aS)-2-pyrazin-2-yl-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (900 mg, 2.64 mmol)in chloroform (10 mL) was added 3-chlorobenzoperoxoic acid (1772 mg,7.91 mmol). The reaction was stirred at 23° C. for 3 h. Additional MCPBA(1.2 g) was added and stirring was continued at room temperature for 18h. The mixture was purified by silica gel column chromatography to givethe title compound (550 mg) as a white solid. LCMS m/z=358.3 [M+H]⁺; ¹HNMR (400 MHz, CDCL₃) δ ppm 0.49 (td, J=4.6, 3.3 Hz, 1H), 1.03 (s, 9H),1.27 (td, J=8.0, 4.9 Hz, 1H), 2.08 (bs, 1H), 2.27-2.33 (m, 1H),2.71-2.76 (m, 1H), 2.93 (d, J=16.8 Hz, 1H), 3.00 (dd, J=16.7, 6.1 Hz,1H), 3.65-3.71 (m, 1H), 3.92-3.97 (m, 2H), 6.97 (d, J=8.5 Hz, 1H), 7.99(dd, J=4.0, 1.4 Hz, 1H), 8.28 (d, J=4.2 Hz, 1H), 8.78 (dd, J=1.4, 0.8Hz, 1H).

A sample was recrystallized from CH₂Cl₂/hexane to give a crystallinesolvate. A thermogravimetric analysis (TGA) thermogram for this solvateshowed a loss of ˜5% weight occurring with a melting endotherm at 164°C.

A non-solvated form of Compound 1 was slurried in CH₂Cl₂ and stirred at˜28° C. overnight. The suspension was filtered using a centrifuge filterand air dried prior to powder X-ray diffraction (PXRD) pattern analysis.The PXRD pattern showed that the material following the CH₂Cl₂ slurrywas indistinguishable from the original solvate form that resulted fromrecrystallization with CH₂Cl₂/hexane. The differential scanningcalorimetry (DSC) thermogram and thermogravimetric analysis (TGA)thermogram for the crystalline CH₂Cl₂ solvate obtained fromrecrystallization using CH₂Cl₂/hexane is shown in FIG. 1; and the PXRDpattern for each of the crystalline CH₂Cl₂ solvates obtained from thetwo different methods (i.e., recrystallization using CH₂Cl₂/hexane; andnon-solvated Compound 1 slurried in CH₂Cl₂) is shown as an overlay inFIG. 2.

Method 2 Step E: Preparation of(1aS,5aS)-2-(Pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicAcid N-oxide

To a suspension of(1aS,5aS)-2-(pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid (68.8 g, 284 mmol) in formic acid (688 mL) was added a 50 wt % aqsolution of hydrogen peroxide (82 mL, 1420 mmol) at room temperature.The mixture was heated to 64° C. The reaction was stirred at 58 to 64°C. for 3 h. Another 8 mL 50% H₂O₂ was added and stirring was continuedanother hour at 60° C. The mixture was let cool to room temperature anddiluted with 1 L water. After storing in an ice bath for 1 h, theprecipitate was collected by filtration, rinsed with water and driedunder vacuum to give the title compound (56.7 g) as a pale yellow solidwhich contains 2% starting material by ¹H NMR. The material wasre-subjected to reaction conditions aforementioned to give the titlecompound (45 g). LCMS m/z=259.2 [M+H]⁺; ¹H NMR (400 MHz, DMSO-d₆) δ 0.42(td, J=4.4, 3.3 Hz, 1H), 1.27 (td, J=7.8, 4.7 Hz, 1H), 2.27-2.33 (m,1H), 2.68-2.73 (m, 1H), 2.75 (dd, J=16.9, 1.5 Hz, 1H), 2.88 (dd, J=16.4,6.4 Hz, 1H), 8.33 (dd, J=4.2, 1.5 Hz, 1H), 8.50 (dd, J=4.2, 0.6 Hz, 1H),8.54 (dd, J=1.5, 0.6 Hz, 1H), 13.08 (s, 1H).

Step F: Preparation of(1aS,5aS)-2-(4-Oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide(Compound 1)

To a suspension of(1aS,5aS)-2-(pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid N-oxide (46.82 g, 181 mmol), (S)-2-amino-3,3-dimethylbutan-1-ol(23.37 g, 199 mmol) and triethylamine (76 mL, 544 mmol) in DMF (400 mL)was added HATU (76 g, 199 mmol). The reaction was stirred at 23° C. for60 min and concentrated. 0.5 M HCl (500 mL) was added. The mixture wasextracted with dichloromethane (3×400 mL). The combined organic extractswere washed with saturated NaHCO₃ (2×250 mL), dried (MgSO₄), filtered,then concentrated to ˜250 mL. To the resulting slurry was added 500 mLof hexanes. The mixture was let stand at room temperature for severalhours and the solid was collected by filtration to give the titlecompound (55 g) as an off-white solid. This material was recrystallizedfrom DCM/hexanes to give the title compound (43.5 g) as a white solid(after drying in vacuum oven at ˜65° C. for 10 days). LCMS m/z=358.3[M+H]⁺; ¹H NMR (400 MHz, CDCl₃) δ ppm 0.49 (td, J=4.6, 3.3 Hz, 1H), 1.03(s, 9H), 1.27 (td, J=8.0, 4.9 Hz, 1H), 2.08 (bs, 1H), 2.27-2.33 (m, 1H),2.71-2.76 (m, 1H), 2.93 (d, J=16.8 Hz, 1H), 3.00 (dd, J=16.7, 6.1 Hz,1H), 3.65-3.71 (m, 1H), 3.92-3.97 (m, 2H), 6.97 (d, J=8.5 Hz, 1H), 7.99(dd, J=4.0, 1.4 Hz, 1H), 8.28 (d, J=4.2 Hz, 1H), 8.78 (dd, J=1.4, 0.8Hz, 1H).

Example 2: Preparation of(1aS,5aS)-2-(4-Oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicAcid ((S)-1-Hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1, DCMSolvate)

The DCM hemi-solvate (10.6% by weight) was obtained by slowcrystallization from CH₂Cl₂ and hexanes and the crystal structure ofthis material was solved, see FIG. 3. Further attempts to isolate thehemi-DCM solvate by forming a slurry of Compound 1 with DCM resulted insubstantially the same DCM Solvates as disclosed in InternationalPublication Number WO2011/025541, an overlay of the PXRDs of theprevious disclosed DCM solvate is shown in FIG. 2.

Example 3: Preparation of(1aS,5aS)-2-(4-Oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicAcid ((S)-1-Hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1,Anhydrous Form)

The anhydrous form of Compound 1 was prepared by recrystallization inDCM and hexanes. The PXRD pattern was characterized, see FIG. 4. Thismaterial melts at ˜162° C. and is a non-solvated form based on TGA FIG.5.

It should be noted that the use of mixtures of DCM/hexanes asrecrystallizing solvents have been observed at different times toprovide different crystal forms.

Example 4: Preparation of(1aS,5aS)-2-(4-Oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicAcid ((S)-1-Hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1,Anhydrous Form) Method 1

To a 4 L reactor equipped with an overhead stirrer, chiller/heater, anda dropping funnel was added(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1, 145 g,406 mmol), acetonitrile (205 mL, 3925 mmol), and water (290 mL). Themixture was heated to 60° C. and then stirred for 60 min. To theresulting reaction was added an additional amount of water (2900 mL),cooled to 0° C., and allowed to stir for 4 h. The mixture was filtered,the solids washed with water and dried under vacuum at 50° C. to provideCompound 1 as the anhydrous form, the material was characterized by PXRD(FIG. 6), and DSC/TGA (FIG. 7).

Method 2

The anhydrous form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1) wasprepared in a similar manner as described in Method 1 except that afterisolating crystalline(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide from theacetonitrile/water crystallizing mixture the material was dried undervacuum at 60° C.±5° C. to provide Compound 1 as the anhydrous form. Theanhydrous form prepared according to Method 2 was characterized by PXRD,DSC, and TGA and was found to be substantially similar to the materialprepared according to Method 1.

Example 5: Preparation of(1aS,5aS)-2-(4-Oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicAcid ((S)-1-Hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1,Acetone Solvate)

The Acetone Solvate of Compound 1 was prepared from a slurry of Compound1 in acetone. The PXRD pattern was characterized, see FIG. 8. Thismaterial showed a loss of weight by TGA of about 5.5%, a desolvationendotherm began at about 100° C. and subsequent melting onset endothermtemperate at about 163° C., see FIG. 9. The acetone solvate wasreproduced from a different lot of anhydrous Compound 1, the PXRD issubstantially identical to that seen in FIG. 8 but with a different lossof acetone as shown by TGA, and thus the stoichiometry of this solvatecan be characterized as a variable or non-stoichiometric acetone solvateof Compound 1.

Example 6: Preparation of(1aS,5aS)-2-(4-Oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicAcid ((S)-1-Hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1,Non-Selective Solvates)

The solvate of Compound 1 was prepared from a slurry of Compound 1 inethyl acetate. The PXRD pattern was characterized, see FIG. 10. Thesubstantially identical PXRD pattern was twice generated from materialprepared by slurring Compound 1 in THF, see FIG. 11. Further, thesubstantially identical PXRD pattern was twice generated from materialprepared by slurring Compound 1 in methyl ethyl ketone (MEK), see FIG.12. Further, the non-selective solvate of Compound 1 prepared usingethyl acetate showed a weight loss of weight of about 4.8% up to about150° C. with an extrapolated onset temperature of 160.8° C.; see TGA andDSC (FIG. 13). The non-selective solvate of Compound 1 prepared usingTHF showed a weight loss of about 6.8% up to about 150° C. with anextrapolated onset temperature of 161.0° C.; see TGA and DSC (FIG. 14).The non-selective solvate of Compound 1 prepared using MEK showed aweight loss of about 4.5% up to about 150° C. with an extrapolated onsettemperature of 160.5° C.; see TGA and DSC (FIG. 15).

The non-selective solvates of Compound 1, independent of the solventused to prepare the solvate, showed substantially the same PXRD as seenin FIGS. 10, 11, and 12 and after desolvation of the solvate each meltswith an extrapolated onset temperature of about 161° C., see FIGS. 13,14, and 15. These solvates appear to be a non-stoichiometric,non-selective solvate crystalline forms of Compound 1 based on TGA.

Example 7: Preparation of(1aS,5aS)-2-(4-Oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicAcid ((S)-1-Hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1, EthylAcetate Solvate)

A ethyl acetate solvate of Compound 1 was prepared by recrystallizationfrom ethyl acetate and heptane.(1aS,5aS)-2-(4-Oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide (Compound 1,430-580 mmol) was dissolved in ethyl acetate (450 mL) at 45° C. Themixture was then cooled to 25° C. and 100 mL of heptane was added to thereactor. The mixture was allowed to stir for 20 min at 22° C. Heptane(2250 mL) was then charged and heated to 55° C. The mixture was allowedto stir overnight at 25° C. The reaction was then cooled to 20° C. andthen filtered and washed with 500 mL heptane. The filter cake was driedunder vacuum at 45° C. The PXRD pattern was characterized, see FIG. 16.The ethyl acetate solvate showed a broad desolvation endotherm fromabout 30-110° C., corresponding to a 1% weight loss on the TGA, see FIG.17. There is a second, larger desolvation/melting endotherm at about115-135° C., corresponding to a TGA weight loss of about 4%. Thecombined weight loss is consistent with the NMR, which showedapproximately 17 mole percent of ethyl acetate, which is equivalent toabout 4.7% by weight. This solvate can contain a trace amount ofheptane.

Example 8: PathHunter β-Arrestin Assay

A: CB₂ Assay

Compound 1 was screened for agonist activity against the human CB₂receptor using the DiscoveRx PathHunter β-arrestin assay which measuresthe β-arrestin binding to the CB₂ receptor upon its activation. CB₂ wascloned into the pCMV-PK vector (DiscoveRx, Fremont, Calif.; catalog#93-0167) and transfected into the CHO-K1 EA-Arrestin parental cell line(DiscoveRx, Fremont, Calif.; catalog #93-0164). CHO-K1 positive clonesstably expressing the CB₂—ProLink fusion protein were identified bytheir responses to the CB₂ agonist CP55,940. Clone #61 was chosen forits big agonist window and homogenous expression as detected by anti-HAflow cytometry

Principle of the Assay:

The PathHunter β-arrestin assay measures the interaction of β-arrestinwith activated GPCRs using Enzyme Fragment Complementation (Yan et al.,J. Biomol. Screen. 7: 451-459, 2002). A small, 42 amino acidβ-galactosidase fragment, Prolink, is fused to the c-terminus of a GPCR,and β-arrestin is fused to the larger β-galactosidase fragment, EA(Enzyme Acceptor). Binding of β-arrestin to the activated GPCR causesthe complementation of the two enzyme fragments, forming an activeβ-galactosidase enzyme which can be measured using the chemiluminiescentPathHunter Flash Detection Kit (DiscoveRx, Fremont, Calif.: catalog#93-0001).

The Assay:

The stable CHO-K1 cells expressing CB₂-Prolink fusion protein wereplated over night in 384-well plates (Optiplate 384-Plus, PerkinElmer,Fremont Calif.; catalog #6007299) at 5000 cells/5 μL/well in theOpti-MEM medium (Invitrogen, Carlsbad, Calif.; catalog #31985088) with1% FBS. 5 uL of test compound diluted in Opti-MEM supplemented with 1%BSA was transferred to each well of the Optiplate. The plates were thenincubated at 37° C./5% CO₂ for two hours. 12 μL of substrate preparedfrom the PathHunter Flash Detection Kit (DiscoveRx, Fremont, Calif.:catalog #93-0001) was transferred to each well of the Optiplate. Theplate was then incubated in the dark at room temperature for 2 h, afterwhich the assay plate was read.

Assay Readout:

β-Arrestin assay readout was accomplished using a PHERAstar (BMG LabtechInc., Durham, N.C.) or an EnVision™ (PerkinElmer, Fremont Calif.)microplate reader.

B: CB₁ Assay

Compound 1 was screened for agonist activity against the human CB₁receptor using the DiscoveRx PathHunter β-arrestin assay which measuresthe β-arrestin binding to the CB₁ receptor upon its activation. CB₁ wascloned into the pCMV-PK vector (DiscoveRx, Fremont, Calif.; catalog#93-0167) and transfected into the CHO-K1 EA-Arrestin parental cell line(DiscoveRx, Fremont, Calif.; catalog #93-0164). CHO-K1 positive clonesstably expressing the CB₁-ProLink fusion protein were identified bytheir responses to the CB₁ agonist CP55,940. Clone #3 was chosen for itsbig agonist window and homogenous expression as detected by anti-HA flowcytometry

Principle of the Assay:

The PathHunter β-arrestin assay measures the interaction of β-arrestinwith activated GPCRs using Enzyme Fragment Complementation (Yan et al.,J. Biomol. Screen. 7: 451-459, 2002). A small, 42 amino acidβ-galactosidase fragment, Prolink, is fused to the c-terminus of a GPCR,and β-arrestin is fused to the larger β-galactosidase fragment, EA(Enzyme Acceptor). Binding of β-arrestin to the activated GPCR causesthe complementation of the two enzyme fragments, forming an activeβ-galactosidase enzyme which can be measured using the chemiluminiescentPathHunter Flash Detection Kit (DiscoveRx, Fremont, Calif.: catalog#93-0001).

The Assay:

The stable CHO-K1 cells expressing CB₁—Prolink fusion protein wereplated over night in 384-well plates (Optiplate 384-Plus, PerkinElmer,Fremont Calif.; catalog #6007299) at 5000 cells/5 μL/well in theOpti-MEM medium (Invitrogen, Carlsbad, Calif.; catalog #31985088) with1% FBS. 5 uL of test compound diluted in Opti-MEM supplemented with 1%BSA was transferred to each well of the Optiplate. The plates were thenincubated at 37° C./5% CO₂ for two h. 12 μL of substrate prepared fromthe PathHunter Flash Detection Kit (DiscoveRx, Fremont, Calif.: catalog#93-0001) was transferred to each well of the Optiplate. The plate wasthen incubated in the dark at room temperature for 2 h, after which theassay plate was read.

Assay Readout:

β-Arrestin assay readout was accomplished using a PHERAstar (BMGLABTECII Inc., Durham, N.C.) or EnVision™ (PerkinElmer, Fremont Calif.)microplate reader.

The EC₅₀ value for hCB₁ was observed to be substantially inactive andthe EC₅₀ value observed for hCB₂ for Compound 1 is shown in thefollowing Table. Compound 1 is a selective agonist for CR₂.

EC₅₀ hCB₂ (nM) Compound 1 5.4

Example 9: Effect of Compound 1 on Osteoarthritis Pain

Injection of monosodium iodoacetate (MIA) into a joint (Kalbhen D. A.,J. Rheumatol., 1987, May; 14 Spec No:130-1; Combe, R., et. al.,Neuroscience Letters, 2004, 370, 236-240) inhibits the activity ofglyceraldehyde-3-phosphate dehydrogenase in chondrocytes, resulting indisruption of glycolysis and eventually in cell death. The progressiveloss of chondrocytes results in histological and morphological changesof the articular cartilage, closely resembling those seen inosteoarthritis patients.

The osteoarthritis was induced in 200 g male Sprague Dawley rats. Afterbrief anaesthesia by isoflurane rats received a single intra-articularinjection of MIA (2 mg) (Sigma Aldrich, Saint Louis, Mo., USA; Cat#19148) dissolved in 0.9% sterile saline in a 50 μL volume administeredthrough the patella ligament into the joint space of the left knee witha 30 G needle. Following the injection, animals were allowed to recoverfrom anaesthesia before being returned to the main housing vivarium.

Typically during disease progression, there was an inflammation periodof 0-7 days post-intra-articular injection followed by progressivedegeneration of the cartilage and subchondral bone from days 14-55.Efficacy studies with a compound of the present invention for paindevelopment took place from day 14 onwards and were performed twice aweek with at least 3 days' wash-out in between each assay. Threedifferent assays were used to measure pain. Tactile allodynia wasmeasured via von Frey assay, hind limb paw weight distribution wasmonitored using an incapacitence tester (Columbus Instruments, Columbus,Ohio, USA) and hind limb grip strength was measured using a gripstrength meter (Columbus Instruments, Columbus, Ohio, USA). Briefly, thevon Frey assay was performed using the standard up down method withvon-Frey filaments. Hind paw weight distribution was determined byplacing rats in a chamber so that each hind paw rests on a separateforce plate of the incapacitence tester. The force exerted by each hindlimb (measured in grams) is averaged over a 3 second period. Threemeasurements were taken for each rat, and the change in hind paw weightdistribution calculated. Peak hind limb grip force was conducted byrecoding the maximum compressive force exerted on the hind limb meshgauge set on the grip strength meter. During the testing, each rat wasrestrained and the paw of the injected knee was allowed to grip themesh. The animal was then pulled in an upward motion until their gripwas broken. Each rat is tested 3 times, with the contralateral paw usedas a control.

Animals were base-lined prior to treatment of the test compound. The MIAtreated groups of rats (6 per group) were then dosed with either vehicle(0.5% methylcellulose, orally), Compound 1 (at 3 mg/kg, 10 mg/kg, and 30mg/kg, orally). Dosing volume was 500 μL. One hour after dosing, vonFrey assay, hind limb weight distribution and/or hind limb grip analysiswas performed to measure the efficacy of the test compound. Increase inpaw withdrawal threshold (PWT) by Compound 1 in comparison with vehicleshown in FIG. 18 was indicative of the test compound exhibitingtherapeutic efficacy in the MIA model of osteoarthritis.

Example 10: Powder X-ray Diffraction

Powder X-ray Diffraction (PXRD) data were collected on an X'Pert PRO MPDpowder diffractometer (PANalytical, Inc.) with a Cu source set at 45 kVand 40 mA, Cu(Kα) radiation and an X'Celerator detector. Samples wereadded to the sample holder and smoothed flat with a spatula and weighpaper. With the samples spinning, X-ray diffractograms were obtained bya 12-min scan over the 2-theta range 5-40 °2θ. Diffraction data wereviewed and analyzed with the X'Pert Data Viewer Software, version 1.0aand X'Pert HighScore Software, version 1.0b.

Example 11: Differential Scanning Calorimetry

Differential scanning calorimetry (DSC) studies were conducted using aTA Instruments, Q2000 at a heating rate 10° C./min. The instruments werecalibrated for temperature and energy using the melting point andenthalpy of fusion of an indium standard. Thermal events (desolvation,melting, etc.) were evaluated using Universal Analysis 2000 software,version 4.1D, Build 4.1.0.16.

Example 12: Thermal Gravimetric Analysis

Thermogravimetric analyses (TGA) were conducted using a TA InstrumentsTGA Q500 or Q5000 at a heating rate 10° C./min. The instruments werecalibrated using a standard weight for the balance, and Alumel andNickel standards for the furnace (Curie point measurements). Thermalevents such as weight-loss are calculated using the Universal Analysis2000 software, version 4.1D, Build 4.1.0.16.

Example 13: Dynamic Moisture-Sorption Analysis

A dynamic moisture-sorption (DMS) study was conducted using a dynamicmoisture-sorption analyzer, VTI Corporation, SGA-100. The instrument wascalibrated using polyvinyl pyrrolidone (PVP) and NaCl. Samples wereprepared for DMS analysis by placing 5 mg to 20 mg of a sample in atared sample holder. The sample was placed on the hang-down wire of theVTI balance. A drying step was run, typically at 40° C. and 0.5-1% RHfor 1 h. The isotherm temperature is 25° C. Defined % RII holdstypically ranged from 10% RII to 90% RII, with intervals of 10 to 20%RH. Λ % weight change smaller than 0.010% over 10 min, or up to 2 h,whichever occurred first, was required before continuing to the next %RH hold. The water content of the sample equilibrated as described abovewas determined at each % RH hold.

The DMS profile (adsorption/desorption isotherm) for the anhydrouscrystalline form of Compound 1 is shown in FIG. 7A. The correspondingdata in tabular form is provided below:

Elapsed Time Weight Weight Sample Sample (min) (mg) (% Change)Temperature RH (%) 46.6 9.6782 0 25.46 1.1 71.4 9.6928 0.151 25.32 29.9491.1 9.7055 0.282 25.31 49.86 111.2 9.7248 0.482 25.3 69.77 129.1 9.73440.581 25.29 79.70 160.1 9.7519 0.762 25.3 89.72 180.1 9.7291 0.526 25.3070.11 200.1 9.7134 0.364 25.3 50.07 218.6 9.6957 0.181 25.29 29.99 234.49.6859 0.080 25.29 10.06

Those skilled in the art will recognize that various modifications,additions, substitutions, and variations to the illustrative examplesset forth herein can be made without departing from the spirit of theinvention and are, therefore, considered within the scope of theinvention.

1. A process for preparing an anhydrous crystalline form of (1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide comprisingcrystallizing(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide from acrystallizing mixture to obtain a crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide in saidcrystallizing mixture, wherein said crystallizing mixture comprisesacetonitrile and water.
 2. The process according to claim 1, whereinsaid crystallizing is conducted at a temperature of about −5° C. toabout 5° C.
 3. The process according to claim 1, wherein saidcrystallizing mixture is prepared by the steps of: 1) dissolving(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide in acetonitrile anda first amount of water to form a first mixture; and 2) adding a secondamount of water to said first mixture to obtain said crystallizingmixture.
 4. The process according to claim 3, wherein said dissolving isconducted at a temperature of about 58° C. to about 62° C.
 5. Theprocess according to claim 3, wherein the molar ratio present in saidfirst mixture between(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide, acetonitrile, andfirst amount water is about 1.0:9.2:37.0 to about 1.0:10.2:41.7.
 6. Theprocess according to claim 3, wherein said adding of said second amountof water to said first mixture is conducted at a rate that thetemperature of the mixture of said second amount of water together withsaid first mixture is at about 25° C. to about 80° C.
 7. The processaccording to claim 3, wherein the molar ratio present in saidcrystallizing mixture between(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide, acetonitrile, andwater in said crystallizing mixture is about 1.0:9.2:414.7 to about1.0:10.2:458.3.
 8. The process according to claim 3, wherein(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide prior to saiddissolving step is selected from the group consisting of: 1) adichloromethane solvate of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide; 2) an acetonesolvate of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide; 3) a non-selectivesolvate of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide; and 4) an ethylacetate solvate of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide; and mixturesthereof. 9-13. (canceled)
 14. The process according to claim 1, whereinafter said crystallizing, said anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide has a chemicalpurity of about 98% or greater.
 15. The process according to claim 1,wherein after said crystallizing, said anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide has an enantiomericexcess of about 98% or greater.
 16. (canceled)
 17. A process forpreparing a pharmaceutical composition comprising the steps of: 1)preparing an anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide according to claim1; and 2) admixing said anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide with apharmaceutically acceptable carrier; wherein said pharmaceuticallyacceptable carrier comprises at least one pharmaceutically acceptableexcipient selected from the group consisting of lactose, mannitol, cornstarch, potato starch, crystalline cellulose, cellulose derivatives,acacia, gelatins, sodium carboxymethyl-cellulose, talc, and magnesiumstearate.
 18. A process for preparing a dosage form comprising the stepsof: 1) preparing an anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide according to claim1; and 2) admixing said anhydrous crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide with apharmaceutically acceptable carrier; wherein said pharmaceuticallyacceptable carrier comprises at least one pharmaceutically acceptableexcipient selected from the group consisting of lactose, mannitol, cornstarch, potato starch, crystalline cellulose, cellulose derivatives,acacia, gelatins, sodium carboxymethyl-cellulose, talc, and magnesiumstearate. 19-27. (canceled)
 28. A pharmaceutical composition comprisingan anhydrous non-solvated crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide and apharmaceutically acceptable carrier, wherein said pharmaceuticallyacceptable carrier comprises at least one pharmaceutically acceptableexcipient selected from the group consisting of lactose, mannitol, cornstarch, potato starch, crystalline cellulose, cellulose derivatives,acacia, gelatins, sodium carboxymethyl-cellulose, talc, and magnesiumstearate.
 29. A unit dosage form comprising an anhydrous non-solvatedcrystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide and apharmaceutically acceptable carrier, wherein said pharmaceuticallyacceptable carrier comprises at least one pharmaceutically acceptableexcipient selected from the group consisting of lactose, mannitol, cornstarch, potato starch, crystalline cellulose, cellulose derivatives,acacia, gelatins, sodium carboxymethyl-cellulose, talc, and magnesiumstearate.
 30. A process of making the pharmaceutical composition ofclaim 28, comprising admixing the anhydrous non-solvated crystallineform of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide with thepharmaceutically acceptable carrier.
 31. A process of making the unitdosage form of claim 29, comprising admixing the anhydrous non-solvatedcrystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide with thepharmaceutically acceptable carrier; and subdividing the admixture intounit dosage forms, each of which comprises a therapeutically effectiveamount of the anhydrous non-solvated crystalline form of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide.
 32. A method forthe treatment of pain in an individual, comprising administering to saidindividual in need thereof, a therapeutically effective amount of apharmaceutical composition according to claim
 28. 33. A method for thetreatment of pain in an individual, comprising administering to saidindividual in need thereof, a therapeutically effective amount of apharmaceutical composition according to claim 28 wherein said pain isassociated with one or more disorders selected from the group consistingof: bone pain, joint pain, muscle pain, dental pain, migraine pain,headache pain, inflammatory pain, neuropathic pain, pain that occurs asan adverse effect of a therapeutic, osteoarthritis pain, cancer pain,multiple sclerosis pain, allergic reactions, nephritic syndrome,scleroderma, thyroiditis, diabetic neuropathy, fibromyalgia, HIVrelated-neuropathy, sciatica, and an autoimmune condition.
 34. A methodfor the treatment of osteoporosis in an individual, comprisingadministering to said individual in need thereof, a therapeuticallyeffective amount of a pharmaceutical composition according to claim 28.35-41. (canceled)
 42. A solvate selected from the group consisting of:an acetone solvate of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide; a non-selectivesolvate of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide; and an ethylacetate solvate of(1aS,5aS)-2-(4-oxy-pyrazin-2-yl)-1a,2,5,5a-tetrahydro-1H-2,3-diaza-cyclopropa[a]pentalene-4-carboxylicacid ((S)-1-hydroxymethyl-2,2-dimethyl-propyl)-amide.
 43. A compositioncomprising a solvate according to claim
 42. 44. A method for thetreatment of pain in an individual, comprising administering to saidindividual in need thereof, a therapeutically effective amount of theunit dosage form according to claim
 29. 45. A method for the treatmentof pain in an individual, comprising administering to said individual inneed thereof, a therapeutically effective amount of the unit dosage formaccording to claim 29; wherein said pain is associated with one or moredisorders selected from the group consisting of: bone pain, joint pain,muscle pain, dental pain, migraine pain, headache pain, inflammatorypain, neuropathic pain, pain that occurs as an adverse effect of atherapeutic, osteoarthritis pain, cancer pain, multiple sclerosis pain,allergic reactions, nephritic syndrome, scleroderma, thyroiditis,diabetic neuropathy, fibromyalgia, HIV related-neuropathy, sciatica, andan autoimmune condition.
 46. A method for the treatment of osteoporosisin an individual, comprising administering to said individual in needthereof, a therapeutically effective amount of the unit dosage formaccording to claim 29.